Objective:To establish a model C-GALAD for detecting hepatocellular carcinoma (HCC) from the chronic liver disease and healthy people based on the serum markers.Methods:A clinical cohort including 229 hepatocellular carcinoma patients, 2 317 patients with chronic liver disease and 982 healthy people, was retrospectively collected from eight hospitals or physical examination institutions from April 2018 to October 2020. The data were divided into a training set and a testing set by stratified sampling with a 6∶4 ratio. A predictive model was established on the training set using a logistic backward regression method and validated on the testing set. In addition, clinical data from March to July 2021 in Beijing You′ an Hospital affiliated to Capital Medical University, including 84 patients with liver cancer and 204 patients with chronic liver disease collected were used for external independent validation of the model. The receiver operating characteristic curve (ROC) area under curve (AUC), the sensitivity and the specificity were used to evaluate the effectiveness of the model.Results:Through the logistic backward regression method, the seven signatures including age, gender, alpha-fetoprotein (AFP), alpha-fetoprotein alloplasm-3 ratio (AFP-L3%), des-gamma-carboxyprothrombin(DCP), platelet (PLT) and total bilirubin (TBIL) were selected as risk factors in the detection model. The area under the ROC curve (AUC) of the model on the testing set was 0.954, with an 88.04% sensitivity and a 94.85% specificity, and the AUC of model on the external independent validation set was 0.943, with an 89.29% sensitivity and a 90.2% specificity, which were better than other published models.Conclusion:The C-GALAD Ⅱ model can accurately predict the risk of hepatocellular carcinoma occurrence, and thus provide a trustworthy diagnosis method of hepatocellular carcinoma.

Objective:To analyze long-term outcomes and prognostic factors of gastric cancer patients after robotic radical total gastrectomy.Methods:A retrospective case-control study was conducted. Inclusion criteria: (1) receiving robotic radical total gastrectomy; (2) gastric adenocarcinoma was confirmed by postoperative pathology; (3) no previous history of other malignant tumors; (4) no preoperative chemotherapy or radiation therapy performed. Exclusion criteria: (1) age <18 years old or age >80 years old;(2)distant metastasis before surgery, or palliative surgery; (3) conversion to laparotomy;(4) R1 or R2 resection; (5)emergency surgery; (6) remnant gastric cancer or recurrence; (7)died of severe complications during hospitalization or within 1 month after surgery. Overall survival rates (OS) and disease-free survival rates (DFS) were evaluated using the Kaplan-Meier method. Cox regression analysis was used to identify prognosis factors for overall survival.Results:According to the above criteria, 166 gastric cancer patients who underwent robotic radical total gastrectomy between March 2010 and November 2018 were included in this study. Roux-en-Y reconstruction was performed in all patients. Reconstruction were achieved using extracorporeal method through a minilaparotomy in 149 case and intracorporeal anastomosis in 17 cases. The number of harvested lymph nodes was (34.8±17.5), and the number of harvested lymph nodes at group 2 was (10.1±6.7). The number of patients with lymph node metastasis of group 2 was 73 (44.0%). The median follow-up time was 25 months (range 2-109). There were 55 (33.1%) cases of recurrence during follow-up. The 3- and 5-year overall survival rates were 55.8% and 46.2% respectively. The 3- and 5-year disease-free survival rates were 53.4% and 45.4% respectively. The 5-year overall survival rates grouped based on TNM stage were 78.9% for stage I, 58.5% for stage II, and 37.1% for stage III. The 5-year disease-free survival rates grouped based on TNM stage were 78.9% for stage I, 59.2% for stage II, and 34.6% for stage III. Univariate analysis suggested that TNM stage, the number of harvested lymph nodes and number of harvested lymph nodes at group 2 were associated with overall survival rates (all P<0.05). Multivariate analyses revealed that TNM stage ( P<0.001; stage IIIB: HR=5.357, 95%CI:1.182 to 24.275; stage IIIC: HR=11.937, 95%CI: 2.677 to 53.226) and number of harvested lymph nodes at group 2 ( P=0.034; 6-10: HR=0.562,95%CI:0.326 to 0.969; >10: HR=0.388, 95%CI: 0.176 to 0.857) were independent prognostic factors for overall survival. Conclusion:The long-term outcomes of robotic radical total gastrectomy were satisfactory. TNM stage and number of harvested lymph nodes at group 2 were independent prognostic factors for overall survival.

Objective:To analyze long-term outcomes and prognostic factors of gastric cancer patients after robotic radical total gastrectomy.Methods:A retrospective case-control study was conducted. Inclusion criteria: (1) receiving robotic radical total gastrectomy; (2) gastric adenocarcinoma was confirmed by postoperative pathology; (3) no previous history of other malignant tumors; (4) no preoperative chemotherapy or radiation therapy performed. Exclusion criteria: (1) age <18 years old or age >80 years old;(2)distant metastasis before surgery, or palliative surgery; (3) conversion to laparotomy;(4) R1 or R2 resection; (5)emergency surgery; (6) remnant gastric cancer or recurrence; (7)died of severe complications during hospitalization or within 1 month after surgery. Overall survival rates (OS) and disease-free survival rates (DFS) were evaluated using the Kaplan-Meier method. Cox regression analysis was used to identify prognosis factors for overall survival.Results:According to the above criteria, 166 gastric cancer patients who underwent robotic radical total gastrectomy between March 2010 and November 2018 were included in this study. Roux-en-Y reconstruction was performed in all patients. Reconstruction were achieved using extracorporeal method through a minilaparotomy in 149 case and intracorporeal anastomosis in 17 cases. The number of harvested lymph nodes was (34.8±17.5), and the number of harvested lymph nodes at group 2 was (10.1±6.7). The number of patients with lymph node metastasis of group 2 was 73 (44.0%). The median follow-up time was 25 months (range 2-109). There were 55 (33.1%) cases of recurrence during follow-up. The 3- and 5-year overall survival rates were 55.8% and 46.2% respectively. The 3- and 5-year disease-free survival rates were 53.4% and 45.4% respectively. The 5-year overall survival rates grouped based on TNM stage were 78.9% for stage I, 58.5% for stage II, and 37.1% for stage III. The 5-year disease-free survival rates grouped based on TNM stage were 78.9% for stage I, 59.2% for stage II, and 34.6% for stage III. Univariate analysis suggested that TNM stage, the number of harvested lymph nodes and number of harvested lymph nodes at group 2 were associated with overall survival rates (all P<0.05). Multivariate analyses revealed that TNM stage ( P<0.001; stage IIIB: HR=5.357, 95%CI:1.182 to 24.275; stage IIIC: HR=11.937, 95%CI: 2.677 to 53.226) and number of harvested lymph nodes at group 2 ( P=0.034; 6-10: HR=0.562,95%CI:0.326 to 0.969; >10: HR=0.388, 95%CI: 0.176 to 0.857) were independent prognostic factors for overall survival. Conclusion:The long-term outcomes of robotic radical total gastrectomy were satisfactory. TNM stage and number of harvested lymph nodes at group 2 were independent prognostic factors for overall survival.

Objective: To analyze the postoperative complications of Da Vinci robotic total gastrectomy for gastric cancer, and explore the risk factors for postoperative complications. Methods: The retrospective case-control study was conducted. The clinicopathological data of 173 patients with gastric cancer who were admitted to the First Affiliated Hospital of Army Medical University from March 2010 to March 2019 were collected. There were 138 males and 35 females, aged from 34 to 76 years, with an average age of 60 years. All the 173 patients underwent Da Vinci robotic total gastrectomy for gastric cancer. Observation indicators: (1) postoperative complications; (2) analysis of risk factors for postoperative complications of Da Vinci robotic total gastrectomy for gastric cancer. Count data were expressed as absolute numbers or percentages. Univariate analysis was performed using the chi-square test or Fisher exact probability. Indicators with P<0.1 were included into multivariate analysis, and multivariate analysis was performed using logistic regression model. Results: (1) Postoperative complications: of the 173 patients, 45 had postoperative complications, with a incidence rate of 26.0%(45/173). Among the 45 patients, 5 had gradeⅠpostoperative complications, 31 had grade Ⅱ postoperative complications, 2 had grade Ⅲa postoperative complications, 3 had grade Ⅲb postoperative complications, 1 had grade Ⅳa postoperative complications, 1 had grade Ⅳb postoperative complications, and 2 had grade Ⅴ postoperative complications. The incidence of serious complications was 5.2%(9/173). Of the 5 patients with gradeⅠcomplications, 1 of fever was improved after antipyretic treatment, 2 of incisional fat liquefaction were improved after dressing change, 1 of vomiting was improved after being given antiemetic, and 1 of delayed recovery of gastrointestinal function was improved after symptomatic treatment. Among 31 patients with gradeⅡcomplications, 12 patients had pulmonary infection, including 6 of pulmonary infection alone, 3 combined with pleural effusion, 1 combined with abdominal infection, 2 combined with intestinal obstruction, and all were improved after conservative treatment; 7 of fever were improved after anti-infection treatment; 4 patients had deep venous catheter infection including 1 combined with bilateral pleural effusion, and were improved after removing catheter and anti-infection treatment; 3 patients had anastomotic leakage including 1 with pulmonary infection and abdominal infection, and were improved after conservative treatment; 2 patients had duodenal stump leakage (1 combined with pulmonary infection, 1 combined with pulmonary infection and pleural effusion) , and were improved after conservative treatment; 1 patient had abdominal hemorrhage, and was improved after conservative treatment; 1 patient had intestinal obstruction, and was improved after conservative treatment; 1 patient had abdominal infection, and was improved after conservative treatment. Of the 2 patients with grade Ⅲa complications, 1 had duodenal stump leakage combined with abdominal abscess, and was improved after puncture and drainage; 1 had pleural effusion combined with pulmonary infection, and was improved after puncture and drainage. Among the 3 patients with grade Ⅲb complications, 1 of abdominal hemorrhage was improved after reoperation, 2 of anastomotic leakage were improved after being placed jejunal nutrition tube under painless gastroscopy. Of the 2 cases, 1 combined with abdominal infection and 1 combined with pleural effusion and abdominal infection were improved after puncture and drainage. Among the 2 patients with grade Ⅳ complications, 1 of Ⅳa encountering respiratory failure was improved after treatment due to misinhalation of anesthesia, and 1 of Ⅳb suffered from multiple organ failure and was improved after treatment due to anastomotic leakage. Two patients with grade V complication died, including one with anastomotic leakage, abdominal hemorrhage, and multiple organ failure, and the other with respiratory failure and cardiac insufficiency. In the 173 patients, the incidence of comprehensive complication index (CCI) ≥ 25.2 was 11.0%(19/173). (2) Analysis of risk factors for postoperative complications of Da Vinci robotic total gastrectomy for gastric cancer: univariate analysis showed that body mass index (BMI), volume of intraoperative blood loss, and operation time were the related factors affecting the postoperative complications (χ²=4.275, 5.057, 5.463, P<0.05). BMI and volume of intraoperative blood loss were the related factors affecting the postoperative serious complications (χ²=7.517, 5.537, P<0.05). Age, BMI and Charlson Comorbidity Index were the related factors affecting CCI ≥25.2 (χ²=8.946, 7.890, 4.062, P<0.05). Multivariate analysis showed that tumor diameter ≥ 3 cm and tumor located at esophagogastric junction were independent risk factors for postoperative complications [odds ratio (OR) =4.350, 2.175, 95% confidence interval (CI): 1.352-14.000, 1.018-4.647, P<0.05)]. BMI ≥25 kg/m² was an independent risk factor for serious complications after operation (OR=5.156, 95%CI: 1.120-23.738, P<0.05). Age ≥60 years, BMI ≥25 kg/m², and history of abdominal surgery were independent risk factors for CCI≥25.2 (OR=30.928, 3.557, 6.009, 95%CI: 1.485-644.19, 1.082-11.691, 1.358-26.592, P<0.05). Conclusions The Clavien-Dindo classification of patients after Da Vinci robotic total gastrectomy for gastric cancer is mostly gradeⅡ. The main complications are pulmonary-related complications. CCI can better predict the risk factors for serious complications after operation. Tumor diameter ≥ 3 cm and tumor located at esophagogastric junction are independent risk factors for postoperative complications; BMI ≥25 kg/m² is an independent risk factor for serious complications; age ≥60 years, BMI ≥25 kg/m², and history of abdominal surgery are independent risk factors for CCI≥25.2.

Objective: To evaluate the clinical application of a novel HIV-1 DNA reagent. Methods: HIV-1-infected and non-infected human blood samples were selected, as well as weakly positive samples, indeterminate samples, specific samples. Compared the result of HIV-1 DNA reagent with HIV-1 infection status (refer to the National Guideline for Detection of HIV/AIDS (2015)), the accuracy of the HIV-1 DNA reagent was evaluated in clinical application; Meanwhile, the commercially available RNA quantification kit was selected as reference reagent for parallel detection, and then the consistency and differences were evaluated between HIV-1 DNA reagent and RNA quantification reagent. Results: A total of 95 whole blood samples were tested by the HIV-1 DNA reagent. Taking the HIV-1 infection status as the reference standard, the result showed that the positive agreement rate was 100% (95%CI: 93.94%-100%), the negative agreement rate was 100% (95%CI: 90.26%-100%), and the overall agreement rate was 100% (95%CI: 96.19%-100%). The Kappa value was 1 (95%CI: 1.00-1.00). The HIV-1 DNA reagent could detect weakly positive samples and indeterminate samples of early infection, and could effectively distinguish false-positive samples tested by the Ag-Ab reagent. The specific samples had no false-positive result . Conclusions The result of HIV-1 DNA reagent were consistent with the HIV-1 infection status. It can be considered as equivalent to the HIV-1 detection reagent commercially available in our country. It can effectively identify the indeterminate samples in the early infection. Compared with the RNA quantification reagent, it can effectively detect HIV-1 DNA of virus reservoirs.

BACKGROUND/AIMS: The Rome III criteria separated chronic constipation into functional constipation (FC) and constipation-predominant irritable bowel syndrome (IBS-C), but some researchers questioned the partitioning and treated both as distinct parts of a continuum. The study aims to explore the similarity and diversity of brain white matter between FC and IBS-C. METHODS: The voxel-wise analysis of the diffusion parameters was used to quantify the white matter changes of female brains in 18 FC patients and 20 IBS-C patients compared with a comparison group with 19 healthy controls by tract-based spatial statistics. The correlations between diffusive parameters and clinical symptoms were evaluated using a Pearson’s correlation. RESULTS: In comparison to healthy controls, FC patients showed a decrease of fractional anisotropy (FA) and an increase of radial diffusivity (RD) in multiple major fibers encompassing the corpus callosum (CC, P = 0.001 at peak), external capsule (P = 0.002 at peak), corona radiata (CR, P = 0.001 at peak), and superior longitudinal fasciculus (SLF, P = 0.002 at peak). In contrast, IBS-C patients showed FA and RD aberrations in the CC (P = 0.048 at peak). Moreover, the direct comparison between FC and IBS-C showed only RD differences in the CR and SLF. In addition, FA and RD in the CC were significantly associated with abdominal pain for all patients, whereas FA in CR (P = 0.016) and SLF (P = 0.040) were significantly associated with the length of time per attempt and incomplete evacuation separately for FC patients. CONCLUSION: These results may improve our understanding of the pathophysiological mechanisms underlying different types of constipation.
Abdominal Pain ; Anisotropy ; Brain ; Constipation ; Corpus Callosum ; Diffusion ; Diffusion Tensor Imaging ; External Capsule ; Female ; Humans ; Irritable Bowel Syndrome ; White Matter

PURPOSE: Cancer-associated fibroblasts (CAFs) activated by cancer cells has a central role in development and malignant biological behavior in colorectal cancer (CRC). Adult fibroblasts do not express Snail, but Snail-positive fibroblasts are discovered in the stroma of malignant CRC and reported to be the key role to chemoresistance. However, the reciprocal effect of CAFs expressed Snail to chemoresistance on CRC cells and the underlying molecular mechanisms are not fully characterized. MATERIALS AND METHODS: Snail-overexpressed 3T3 stable cell lines were generated by lipidosome and CT26 mixed with 3T3-Snail subcutaneous transplanted CRC models were established by subcutaneous injection. Cell Counting Kit-8, flow cytometry and western blotting assays were performed, and immunohistochemistry staining was studied. The cytokines participated in chemoresistance was validated with reverse transcriptase-polymerase chain reaction and heatmap. RESULTS: Snail-expression fibroblasts are discovered in human and mouse spontaneous CRCs. Overexpression of Snail induces 3T3 fibroblasts transdifferentiation to CAFs. CT26 co-cultured with 3T3-Snail resisted the impairment from 5-fluorouracil and paclitaxel in vitro. The subcutaneous transplanted tumor models included 3T3-Snail cells develop without restrictions even after treating with 5-fluorouracil or paclitaxel. Moreover, these chemoresistant processes may be mediated by CCL1 secreted by Snail-expression fibroblasts via transforming growth factor β/nuclear factor-κB signaling pathways. CONCLUSION: Taken together, Snail-expressing 3T3 fibroblasts display CAFs properties that support 5-fluorouracil and paclitaxel chemoresistance in CRC via participation of CCL1 and suggest that inhibition of the Snail-expression fibroblasts in tumor may be a useful strategy to limit chemoresistance.
Adult ; Animals ; Blotting, Western ; Cell Count ; Cell Line ; Colorectal Neoplasms* ; Cytokines ; Drug Resistance, Multiple ; Fibroblasts* ; Flow Cytometry ; Fluorouracil ; Humans ; Immunohistochemistry ; In Vitro Techniques ; Injections, Subcutaneous ; Mice ; Paclitaxel ; Snails ; Transforming Growth Factors

BACKGROUND:Our previous studies have found that polygonatum sibiricum polysaccharide (PSP) promotes osteogenic differentiation of bone marrow mesenchymal stem cel s (BMSCs) by Wnt/β-catenin signaling pathway, but the molecular mechanism is unclear.OBJECTIVE:To investigate the effect of PSP promoting the osteogenic differentiation via Wnt signaling pathways in BMSCs after LRP5 silencing. METHODS:LRP5 interference vectors were constructed and then transfected into C57BL/6 mouse BMSCs cultured in vitro. The transfection efficiency of cel s was calculated under fluorescence inverted microscope and the expression of LRP5 protein was detected by western blot assay. The osteogenic potential of BMSCs after LRP5-siRNA transfection was analyzed by alkaline phosphatase staining, alizarin red staining and western blot assay. Effect of PSP on the osteogenic differentiation of LIRP5-silenced mouse BMSCs was detected by real-time PCR and dual luciferase assay. RESULTS AND CONCLUSION:Compared with the control group, the mineralization ability, the mRNA expressions of Runx2 and Osterix, and the protein expression of LRP5 were significantly decreased in the LRP5-siRNA group (P<0.05). PSP could promote LRP5-siRNA transfected mouse BMSCs differentiating into osteoblasts and significantly upregulated the expressions ofβ-catenin and Osterixin, and also induced the high expression of luciferase reporter gene (TOPFlash) containing wild type TCF binding sites (P<0.05). To conclude, LRP5 plays an important role in the process of mouse BMSCs differentiating into osteoblasts. PSP can promote the osteogenic differentiation of mouse BMSCs by activating the Wnt/β-catenin signaling pathway independent on LRP5.

BACKGROUND: Bone marrow-derived mononuclear cells (BM-MNCs) hold the potential of differentiating into osteoclasts. Polygonatum sibiricum polysaccharide (PSP) may inhibit the differentiation of BM-MNCs into osteoclasts and it is expected to become a new drug for the treatment of osteoporosis. OBJECTIVE: To investigate the effect of PSP on the differentiation of mouse BM-MNCs into osteoclasts induced by receptor activator of nuclear factor kappa-B ligand (RANKL) and bone resorption in vivo. METHODS: Mouse bone marrow-derived macrophages cultured in vitro, the effect of macrophage colony stimulating factor and PSP (5, 10, 20, 40, 80,160, 320, 640, 1280, 2560 mg/L) on the proliferation of mouse BM-MNCs was detected by cell counting kit-8 assay to determine the PSP concentration range; the mouse BMMs were cultured and induced in DMEM medium containing macrophage colony stimulating factor, RANKL and 5, 10, 20, 40, 80,160, 320, 640 mg/L PSP, respectively; those cultured without PSP served as control group. The morphological changes of cells were observed under an inverted microscope.; the number of osteoclasts was detected by tartrate-resistant acid phosphatase staining; the mRNA expression levels of osteoclast-related genes including tartrate-resistant acid phosphatase, matrix metalloproteinase-9, cathepsin K, and nuclear factor of activated T cells c1 were evaluated by quantitative real-time PCR. A mouse model of calvarial osteolysis induced by lipopolysaccharide was established to receive PSP intervention, and then micro CT scanning, three-dimensional reconstruction and relevants software were used for quantitative analysis of bone volume/volume percentage, trabecular number, trabecular bone spacing and thickness. The number of osteoclasts was identified by tartrate-resistant acid phosphatase staining and quantitative analysis of bone resorption area was conducted. RESULTS AND CONCLUSION: Compared with the control group, the concentration of PSP below 640 mg/L showed no significant effect on the proliferation of BMMs (P > 0.05). Different concentrations of PSP (40-640 mg/L) significantly reduced the number of osteoclasts, osteoclast differentiation and maturation, and the mRNA expression levels of tartrate-resistant acid phosphatase, matrix metalloproteinase-9, cathepsin K, and nuclear factor of activated T cells c1 TRAP, MMP-9, CtsK and NFATc1 (P < 0.05). Compared with lipopolysaccharide, PSP could effectively alleviate the lipopolysaccharide-induced calvarial osteolysis, and the bone volume/volume percentage, trabecular number, and trabecular bone spacing were significantly decreased (P < 0.05); additionally, the number of osteoclasts and the area of bone resorption were decreased significantly (P < 0.01). To conclude, PSP can inhibit the differentiation and maturation of mouse BMMs to osteoclasts and alleviate lipopolysaccharide-induced calvarial osteolysis.

BACKGROUND:Posterior lamina resection often causes loss of spinal stability, so screw rod internal fixation technology is needed to maintain the stability of lumbar spine. Finite element analysis can be used to simulate the stress distribution of the spine and internal fixation system after spinal surgery. OBJECTIVE: To build three-dimensional finite element model of spinal L1 to L3, analyze the spinal stability and stress distribution after the total laminectomy and insertion of bilateral pedicle screw using finite element method. METHODS: L1-L3 CT data could be colected from an adult healthy male volunteer. Mimics14.01, 3-matic(V6.0) and Ansys 15.0 could be used to set up the intact lumbar spine finite element model of L1-L3 (group A), the L1-L3 finite element model after L2 total laminectomy (group B), and the finite element model of L2 total laminectomy and insertion of bilateral pedicle screw (group C). We used software to simulate flexion, extension, lateral bending and axial rotation, and three kinds of models received finite element analysis. RESULTS AND CONCLUSION: (1) Based on the maximum of Von Mises under different motion states, the maximum stress was significantly lower in group A than in group B (P< 0.05). The maximum stress was significantly lower in group B than in group C (P < 0.05). (2) Based on the total deformation under different motion states, the total deformation was significantly lower in group A than in group B (P < 0.05). The total deformation was significantly lower in group C than in groups A and B (P < 0.05). (3) After the total laminectomy, vertebral body stress increased, especialy in the lamina, pedicle and joints. The range of motion of the vertebral body increased, which influenced the stability of the vertebral body. Internal fixation could decrease range of motion. Stress concentrated on the screw. Stress on the vertebral plate and pedicle decreased. The stability of vertebral body increased. Excessive stress concentrated on screw system wil increase the risk of screw breakage.