ABSTRACT
@#Resection is one of the most important treatments for esophageal squamous cell carcinoma, and routine postoperative follow-up is an effective method for early detection and treatment of recurrent metastases, which can improve patients' quality of life and prognosis. This consensus aims to provide a reference for colleagues responsible for postoperative follow-up of esophageal squamous cell carcinoma patients in China, and further improve the standardization of the diagnosis and treatment of esophageal squamous cell carcinoma.
ABSTRACT
Objective:To investigate the effects of human umbilical cord-derived mesenchymal stem cells (hUC-MSC) and their conditioned medium on proliferation, migration and apoptosis of human non-small cell lung cancer (NSCLC) polyploid A549 cells.Methods:A549 cells in logarithmic phase were selected. After induction treatment with 1 μmol/L docetaxel for 24 h, DMEM/F12 medium with 10% fetal bovine serum was used to culture the cells for 3 d, finally the polyploid A549 cells model was successfully established. After finishing the separation and culture of hUC-MSC, hUC-MSC conditioned medium was prepared. Normally cultured polyploid A549 cells were treated as the control group, conditioned medium cultured polyploid A549 cells were treated as the conditioned medium group. hUC-MSC was co-cultured with polyploid A549 cells, and the ratio of the total number of cells was 2:1 and 5:1, respectively, which were recorded as MSC 1 group and MSC 2 group. Cells in each group were continually cultured for 48 h or 72 h. Proliferation and apoptosis of polyploid A549 cells in each group were detected by using flow cytometry, cell migration ability was detected by using Transwell assay, and the expressions of migration and apoptosis-related proteins were detected by using Western blotting.Results:Polyploid A549 cells model was successfully established and hUC-MSC was cultured separately. The result of cell proliferation detected by flow cytometry showed that at 48 h, the mean fluorescence intensity of the control group, conditioned medium group, MSC 1 group and MSC 2 group was 1 695±305, 2 020±85, 1 259±35 and 1 356±33, respectively, and the difference was statistically significant ( F = 14.00, P < 0.05); at 72 h, the mean fluorescence intensity of the control group, conditioned medium group, MSC 1 group and MSC 2 group was 1 052±77, 1 309±24, 864±201 and 1 103±237, respectively, and the difference was statistically significant ( F = 3.90, P > 0.05). The result of Transwell assay showed that at 48 h, the number of cell migration in the control group, conditioned medium group, MSC 1 group and MSC 2 group was 52±9, 57±12, 68±8 and 75±11, respectively, and the difference was statistically significant( F = 32.16, P < 0.05); the number of cell migration in each experimental group was all higher than that in the control group (all P < 0.05). The percentage of apoptotic cells in the control group, conditioned medium group, MSC 1 group and MSC 2 group was (15.53±4.27)%, (13.77±1.75)%, (3.60±0.50)% and (2.33±0.06)%, respectively, and the difference was statistically significant ( F = 182.36, P < 0.05); there was no statistically significant difference between the control group and conditioned medium group ( P > 0.05); there were statistically significant differences between MSC 1 group and the control group, MSC 2 group and the control group (both P < 0.05). Western blotting results showed that compared with the control group, the expression of migration-related protein matrix metallopeptidase 9 (MMP-9) was increased, the expression of pro-apoptotic protein bax was reduced, the expression of anti-apoptotic protein bcl-xL was increased in conditioned medium group, MSC 1 group and MSC 2 group. Conclusions:hUC-MSC can improve the migration and anti-apoptotic ability of polyploid A549 cells, suggesting that hUC-MSC may affect the survival of tumor cells during the process of chemotherapy damage and repair.
ABSTRACT
Anastomotic leakage (AL) has always been a persistent issue for colorectal surgeons. It is still difficult to reduce the incidence of AL despite the advances in technology and equipment. With the development of evidence-based medicine, increasing high-risk factors for AL have been identified. How to efficiently and systematically combine and quantify these isolated risk factors to provide a scientific early warning of AL in clinical practices and help surgeons in choosing the optimal prophylactic strategies, is of great significance for reducing the incidence of AL. There are generally two types of AL prediction models in colorectal surgery, including prognostic models (for preoperative and intraoperative AL prediction) and diagnostic models (for early warning and improving the early diagnosis rate of AL). Prophylactic strategies for AL include stabilizing the underlying diseases, improving anemia and hypoalbuminemia, choosing an appropriate operative time window, and emphasizing and improving anastomotic techniques (including choosing an appropriate size of stapler). However, a prophylactic ostomy is still the most common method for surgeons. However, how to reduce the morbidity of complications following prophylactic ostomy and how to avoid the conversion of the prophylactic stoma to permanent stoma need further study.
Subject(s)
Humans , Anastomotic Leak/etiology , Colorectal Surgery/adverse effects , Digestive System Surgical Procedures/adverse effects , Anastomosis, Surgical/methods , Risk FactorsABSTRACT
Babesia microti is one of the most common causative agents of babesiosis. A sensitive and rapid detection is necessary for screening potentially infected individuals. In this study, B. microti cytochrome c oxidase subunit I (cox1) was selected as the target gene, multiple primers were designed, and optimized by a recombinase-aided amplification (RAA) assay. The optimal primers and probe were labeled with fluorescein. The sensitivity of fluorescent RAA (fRAA) was evaluated using gradient diluents of the cox1 recombinant plasmid and genomic DNA extracted from whole blood of B. microti infected mice. The specificity of fRAA was assessed by other transfusion transmitted parasites. The analytical sensitivity of the fRAA assay was 10 copies of recombinant plasmid per reaction and 10 fg/µl B. microti genomic DNA. No cross-reaction with any other blood-transmitted parasites was observed. Our results demonstrated that the fRAA assay would be rapid, sensitive, and specific for the detection of B. microti.
ABSTRACT
Hepatic encephalopathy (HE) is a common serious complication of liver cirrhosis, with sudden onset, indicating a poor prognosis in patients with chronic liver disease. Minimal hepatic encephalopathy (MHE) is an early stage of HE with no apparent symptoms, but it shows abnormal results in neuropsychological and/or neurophysiological tests. MHE affects patients' quality of life, employability, driving ability, and has a high risk of developing overt hepatic encephalopathy (OHE). This article aims to explore various diagnostic methods, strengthen the routine work of clinicians in diagnosis and treatment, and develop an effective MHE screening protocol.
Subject(s)
Hepatic Encephalopathy/diagnosis , Humans , Liver Cirrhosis/diagnosis , Liver Diseases , Mass Screening , Neuropsychological Tests , Psychometrics , Quality of LifeABSTRACT
Objective To develop a fluorescent recombinase-aided isothermal amplification (RAA)-based nucleic acid assay for detection of Leshimania. Methods Specific primers and probes were designed targeting Leishmania internal transcribed spacer 1 (ITS1) gene for RAA assay, and a fluorescent RAA assay was developed for detection of Leishmania following screening of primer pairs and optimization of primer and probe concentrations. The sensitivity of RAA assay for detection of Leishmania was evaluated using recombinant plasmid containing Leishmania ITS1 gene sequences at different copies and Leshimania genomic DNA at different concentrations as templates, and the specificity of RAA assay for detection of Leishmania was evaluated using the genomic DNA of transfusion-transmitted parasites, including Babesia microti, Toxoplasma gondii, Plamodium vivax, P. ovale, P. falciparum, P. malariae, L. donovani and L. infantum. Results After the optimal primer pair was screened from 9 pairs of primer combinations, the final primer and probe concentrations were optimized as 0.3 μmol/L and 0.08 μmol/L, respectively. Nucleic acid detection of Leishmania was completed by the fluorescent RAA assay at an isothermal temperature of 39 °C within 20 min. Remarkable florescent signals were seen within 5 min following RAA detection of genomic DNA of L. donovani and L. infantum, and no cross-reactions were observed with B. microti, T. gondii, P. vivax, P. ovale, P. falciparum or P. malariae. The lowest limitation of detection of the fluorescent RAA assay was 10 copies/μL recombinant plasmid containing Leishmania ITS1 gene sequences and 1 fg/μL Leishmania genomic DNA. Conclusions A rapid, simple, sensitive and specific fluorescent RAA assay is successfully developed for detection of L. donovani and L. infantum, which is effective for field screening of leishmaniasis.
ABSTRACT
Objective To evaluate the efficiency of a recombinase-aided amplification (RAA) assay for the detection of Schistosoma japonicum infections in Oncomelania hupensis snails. Methods A group test was employed. Fifty Oncomelania snails were collected as a detection sample. The detection samples without infected snails were designated as negative specimens, while the detection samples that contained different numbers of infected snails were designated as positive specimens. A total of 10 negative specimens, 10 positive specimens containing 1 infected snail, 20 positive specimens containing 2 infected snails and 10 positive specimens containing 3 infected snails were assigned. Following random grouping, 40 specimens were subject to the florescent RAA assay using a blind method. The miradium shedding method served as a gold standard, and the sensitivity, specificity, Youden’s index and coincidence rate of the florescent RAA assay were estimated. In addition, 20 samples consisted of 5 negative specimens and 15 positive specimens with 1, 2 and 3 infected snails respectively were grouped randomly. The same specimens were detected using the crushing method and fluorescent RAA assay with the blind method in a paired-design manner. Then, the test results were compared and analyzed. Results Florescent RAA assay detected 29 positives in the 30 specimens containing different numbers of infected snails, with a sensitivity of 96.67%, and 8 negatives in the 10 detection specimens without infected snails, with a specificity of 80.00%, showing a Youden’s index of 0.77. The coincidence rate was 100% among 10 repeated assays for a detection specimen. In addition, there was no significant difference in the detection of infected snails between the florescent RAA assay and the crushing method (χ2 = 0, P > 0.05), and the actual coincidence rates of the florescent RAA assay and crushing method were 95.00% (19/20) and 90.00% (18/20) with the real results, respectively. Conclusion Fluorescent RAA assay has a favorable efficiency for the detection of S. japonicum infections in Oncomelania snails, which shows a potential in screening of S. japonicum-infected Oncomelania snails.
ABSTRACT
Anoctamin 1 (ANO1) is a kind of calcium-activated chloride channel involved in nerve depolarization. ANO1 inhibitors display significant analgesic activity by the local peripheral and intrathecal administration. In this study, several thiophenecarboxylic acid and benzoic acid derivatives were identified as novel ANO1 inhibitors through the shape-based virtual screening, among which the 4-arylthiophene-3-carboxylic acid analogues with the best ANO1 inhibitory activity were designed, synthesized and compound
ABSTRACT
Objective To develop a rapid test for detection of Schistosoma japonicum specific gene fragments based on the recombinase-aided isothermal amplification assay (RAA) and nucleic acid dipstick test. Methods The S. japonicum SjG28 gene fragment was selected as the target gene fragment, and the primers and fluorescent probe were designed and synthesized. Then, a S. japonicum nucleic acid dipstick test was established. The sensitivity of this dipstick test was evaluated by detecting different copies of recombinant plasmids containing the S. japonicum SjG28 gene fragment and different concentrations of genomic DNA from adult worms of S. japonicum, and the specificity of the dipstick test was evaluated by detecting the genomic DNA from Clonorchis sinensis, S. mansoni, Ancylostoma duodenale, S. haematobium, Babesia and Paragonimus westermani. Results The S. japonicum nucleic acid dipstick test based on the S. japonicum SjG28 gene fragment showed the minimum detectable limit of 10 copies/μL of the recombinant plasmid containing the S. japonicum SjG28 gene fragment and the minimum detectable limit of 1 pg/μL of S. japonicum genomic DNA, and the dipstick assay tested negative for the genomic DNA from C. sinensis, S. mansoni, A. duodenale, S. haematobium, Babesia and P. westermani. Conclusion A rapid, simple, and visualized assay is established for detection of S. japonicum specific gene fragments based on RAA and nucleic acid dipstick test.
ABSTRACT
Objective:To investigate the clinical efficacy of Da Vinci robotic assisted and laparos-copic assisted complete mesocolic excision (CME) for right hemicolon cancer.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopatho-logical data of 119 patients with right hemicolon cancer who were admitted to Daping Hospital, Army Medical University from July 2016 to July 2019 were collected. There were 63 males and 56 females, aged (61±11)years. All the 119 patients underwent CME of right hemicolon. Of 119 patients, 37 cases undergoing Da Vinci robotic assisted CME of right hemicolon were divided into robotic group and 82 cases undergoing laparoscopic assisted CME of right hemicolon were divided into laparoscopic group. Observation indicators: (1) the propensity score matching conditions and comparison of general data between the two groups after propensity score matching; (2)intraoperative and postoperative situations; (3) postoperative pathological examination; (4)follow-up. Follow-up was conducted by outpatient examination or telephone interview to detect tumor metastasis and survival of patients after surgery up to August 2019. The propensity score matching was conducted by 1∶1 matching using the nearest neighbor method. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Count data were represented as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. The Kaplan-Meier method was used to calculate survival rate and the GraphPad Prism 5 software was used to draw survival curve. The Log-rank test was used for survival analysis. Results:(1) The propensity score matching conditions and comparison of general data between the two groups after propensity score matching: 68 of 119 patients had successful matching, including 34 cases in each group. Before propensity score matching, cases undergoing surgery by surgeon A or surgeon B were 32, 5 of the robotic group, versus 49, 33 of the laparoscopic group, showing a significant difference between the two groups ( χ2=8.381, P<0.05). After propensity score matching, the gender (males or females), age, body mass index (BMI), cases with tumor classified as stageⅠ, stage Ⅱ or stage Ⅲ of TNM staging, cases with tumor located at ileocecal region, ascending colon, hepatic flexor of colon or transverse colon, cases undergoing surgery by surgeon A or surgeon B were 17, 17, (62±10)years, (22.4±2.7)kg/m 2, 4, 14, 16, 3, 15, 10, 6, 29, 5 of the robotic group, versus 15, 19, (62±11)years, (22.4±2.8)kg/m 2, 4, 18, 12, 2, 19, 7, 6, 30, 4 of the laparoscopic group, showing no significant difference between the two groups ( χ2=0.236, t=0.127, 0.044, χ2=1.071, 1.200, 0.000, P>0.05). (2) Intraoperative and postoperative situations: after propensity score matching, the operation time, volume of intraoperative blood loss, cases undergoing conversion to open surgery, time to postoperative initial out-of-bed activities, time to postoperative first flatus, time to postoperative initial liquid food intake, duration of postoperative hospital stay and treatment expenses were (235±50)minutes, (73±45)mL, 0, (1.9±0.7)days, (2.9±1.2)days, (3.1±2.4)days, (9.1±4.9)days, (9.6±1.8)×10 4 yuan of the robotic group, versus (183±35)minutes, (74±74)mL, 1, (2.1±0.6)days, (3.3±1.4)days, (3.5±4.2)days, (9.1±3.9)days, (6.3±1.6)×10 4 yuan of the laparoscopic group, respectively. There were significant differences in the operation time and treatment expenses between the two groups ( t=5.050, 8.165, P<0.05) while there was no significant difference in the volume of intraoperative blood loss, time to postoperative initial out-of-bed activities, time to postoperative first flatus, time to postoperative initial liquid food intake or duration of postoperative hospital stay between the two groups ( t=0.118, ?0.462, ?1.129, ?1.291, 0.027, P>0.05). There was no significant difference in the conversion to open surgery between the two groups ( P>0.05). Five patients of the robotic group and 7 patients of the laparoscopic group had postoperative complications. There was no significant difference in the postoperative complications between the two groups ( χ2=0.405, P>0.05). (3) Postoperative pathological examination: after propensity score matching, cases with R 0 resection, the number of lymph node dissected, cases with lymph node metastasis and cases with tumor differentiation as well differentiated adenocarcinoma, moderately differentiated adeno-carcinoma, poorly differentiated adenocarcinoma or mucinous adenocarcinoma were 34, 17±5, 14, 1, 22, 6, 5 of the robotic group, versus 34, 17±5, 12, 2,20, 2, 10 of the laparoscopic group, respectively. There was no significant difference in the R 0 resection between the two groups ( P>0.05) and there was no significant difference in the number of lymph node dissected, lymph node metastasis and tumor differentiation between the two groups ( t=0.488, χ2=0.249, 4.095, P>0.05). (4) Follow-up: after propensity score matching, 68 patients were followed up for 1?36 months, with a median follow-up time of 24 months. The follow-up time was (20±13)months of the robotic group, versus (21±13)months of the laparoscopic group, showing no significant difference between the two groups ( t=0.409, P>0.05). During the follow-up, 3 cases of the robotic group and 4 cases of the laparoscopic group had tumor distant metastasis. The disease-free survival rate and overall survival rate at postoperative 3 years were 83.9% and 86.8% of the robotic group, versus 82.0% and 86.6% of the laparoscopic group, showing no significant difference between the two groups ( χ2=0.188, 0.193, P>0.05). Conclusion:Da Vinci robotic assisted CME for right hemicolon cancer is safe and feasible.
ABSTRACT
Objective:To investigate the influencing factors for celiac lymph node metastasis in thoracic esophageal squamous cell carcinoma (TE-SCC), construct a prediction model of celiac lymph node metastasis in TE-SCC, and stratify the probability of celiac lymph node metastasis.Methods:The retrospective case-control study was conducted. The clinicopathological data of 443 patients with TE-SCC who underwent thoracoscopic and laparoscopic esophagectomy with systematic lymph node dissection in the First Affiliated Hospital of Zhengzhou University between March 2015 and April 2019 were collected. There were 259 males and 184 females, aged from 41 to 81 years, with a median age of 64 years. The nomogram prediction model was constructed based on the results of multivariate analysis of influencing factors for celiac lymph node metastasis in TE-SCC, of which calibration curve and decision curve were drawed. The predictive performance was evaluated using the concordance index. The score for celiac lymph node metastasis in TE-SCC predicted by nomogram model was used for further recursive partitioning analysis, and patients were stratified into risk subgroups using the decision-making tree model. Observation indicators: (1) celiac lymph node metastasis in TE-SCC; (2) analysis of influencing factors for celiac lymph node metastasis in TE-SCC; (3) construction of nomogram prediction model of celiac lymph node metastasis in TE-SCC; (4) construction of decision-making tree model of celiac lymph node metastasis in TE-SCC and risk subgroup analysis of celiac lymph node metastasis probability. Measurement data with skewed distribution were represented as M (range). Count data were represented as absolute numbers and percentages, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data between groups was analyzed using the nonparametric rank sum test. Multivariate analysis was performed using the Logistic regression model. Based on Logistic regression model multivariate analysis, a new nomogram model was constructed using the RStudio 3.4 software. Results:(1) Celiac lymph node metastasis in TE-SCC: celiac lymph node metastasis was found in 89 of the 443 patients, with a celiac lymph node metastasis rate of 20.09%(89/443). (2) Analysis of influencing factors for celiac lymph node metastasis in TE-SCC. Results of univariate analysis showed that tumor location, tumor length, tumor differentiation degree, pathological T staging, nerve invasion, vessel invasion, and thoracic lymph node metastasis were related factors for celiac lymph node metastasis in TE-SCC ( χ2=12.177, Z=-2.754, -4.218, -4.254, χ2=3.908, 33.025, 30.387, P<0.05). Results of multivariate analysis showed that tumor location, vessel invasion, and thoracic lymph node metastasis were independent influencing factors for celiac lymph node metastasis in TE-SCC ( odds ratio=2.165, 3.442, 2.876, 95% confidence interval: 1.380-3.396, 1.787-6.633, 1.631-5.071, P<0.05). (3) Construction of nomogram prediction model of celiac lymph node metastasis in TE-SCC: based on the factors screened by multivariate analysis, including tumor location, vessel invasion, and thoracic lymph node metastasis, the nomogram prediction model of celiac lymph node metastasis in TE-SCC was established, with the concordance index of 0.846. The calibration curve showed a high consistency between the celiac lymph node metastasis probability estimated by the prediction model and the actual rate of celiac lymph node metastasis. The decision curve showed that the nomogram prediction model of celiac lymph node metastasis in TE-SCC had a good prediction value when the probability threshold was 0.001-0.819.(4) Construction of decision-making tree model of celiac lymph node metastasis in TE-SCC and risk subgroup analysis of celiac lymph node metastasis probability: patients were stratified into six risk subgroups using the decision-making tree model based on the celiac lymph node metastasis probability. The group A included patients with no vessel invasion+negative thoracic lymph node, group B included patients with no vessel invasion+the number of positive thoracic lymph nodes of 1-3, group C included patients with no vessel invasion+the number of positive thoracic lymph nodes of ≥4, group D included patients with vessel invasion+the number of positive thoracic lymph nodes of 0-2+upper or middle thoracic esophageal carcinoma, group E included patients with vessel invasion+the number of positive thoracic lymph nodes of 0-2+lower thoracic esophageal carcinoma, group F included patients with vessel invasion+the number of positive thoracic lymph nodes of ≥3. The group A was low-risk group with the celiac lymph node metastasis probability of 11%, group B and D were intermediate low-risk groups with the celiac lymph node metastasis probability of 27% and 21%, group C and E were the intermediate high-risk groups with the celiac lymph node metastasis probability of 56% and 55%, and group F was high-risk group with the celiac lymph node metastasis probability of 80%. Conclusions:The tumor location, vessel invasion, and thoracic lymph node metastasis are independent influencing factors for celiac lymph node metastasis in TE-SCC. Vessel invasion has the dominant influence on celiac lymph node metastasis, followed by the number of positive thoracic lymph nodes, and then the tumor location. Patients can be stratified into six risk subgroups based on the nomogram prediction model and decision-making tree model of celiac lymph node metastasis in TE-SCC.
ABSTRACT
Objective:To study the migration of polyploid tumor cells induced by docetaxel, the characteristics of epithelial-mesenchymal transition, and the killing effect of immune cells on them, in order to explore the potential role of polyploid tumor cells in tumor recurrence.Methods:The human non-small cell lung cancer A549 cells were treated with 1 μmol/L docetaxel for 24 h, and the cells were collected as Doc 1 d group. After drug removal, the cells were cultured in fresh and complete medium for 3 or 5 days, then the cells were collected as Doc 3 d group or Doc 5 d group respectively. The A549 cells were treated with DMSO for 24 h as control group. Immunofluorescence staining was used to detect cell morphology, flow cytometry was used to analyze cell ploidy, scratch test was used to detect cell migration, Western blotting was used to detect the expression of epithelial-mesenchymal transition related proteins, and lactate dehydrogenase release method was used to evaluate the killing activity of cytokine-induced killer (CIK) cells.Results:Compared with the control group, most of the cells in the Doc 1 d group, Doc 3 d group and Doc 5 d group were apoptotic, a few of the surviving cells were significantly larger, and the nucleus was polynuclear. The proportions of polyploid cell subset (DNA content > 4N) in the control group, Doc 1 d group, Doc 3 d group and Doc 5 d group were (1.93±0.55)%, (22.97±2.37)%, (51.30±12.51)% and (67.87±8.31)% respectively, and the difference among the four groups was statistically significant ( F=26.521, P<0.001). The proportion of polyploid cell subset in Doc 1 d group, Doc 3 d group and Doc 5 d group was significantly higher than that in the control group (all P<0.001). With the prolongation of withdrawal time, the proportion of polyploid cell subset in Doc 3 d group and Doc 5 d group was significantly higher than that in Doc 1 d group ( P=0.009; P=0.004). After 24 h and 48 h culture, the wound healing rates of the control group were both 100%, and the wound healing rates of the Doc 3 d group were (39.10±2.12)% and (46.13±5.32)% respectively, with no significant difference ( t=2.126, P=0.051). Compared with the control group at 24 h and 48 h, the cell migration abilities of Doc 3 d group were significantly lower ( t=49.756, P<0.001; t=30.825, P<0.001). Compared with the control group, the expression of E-cadherin protein decreased gradually in the Doc 1 d group, Doc 3 d group and Doc 5 d group, the expression of Vimentin protein increased gradually, and the expressions of Snail protein and N-cadherin protein did not change significantly. The killing efficiencies of CIK cells against the cells of the control group, Doc 3 d group and Doc 5 d group were (27.27±1.91)%, (17.87±2.35)%, (9.47±0.51)% respectively, and the difference was statistically significant ( F=11.294, P<0.001). The killing efficiency of Doc 3 d group and Doc 5 d group was significantly lower than that of the control group ( P=0.004; P<0.001). The killing efficiency of Doc 5 d group was significantly lower than that of Doc 3 d group ( P=0.003). Conclusion:The migration ability of polyploid tumor cells induced by docetaxel is weakened, but epithelial-mesenchymal transition is likely to occur, and the killing effect of immune cells on them is reduced.
ABSTRACT
Objective To compare the changes in cardiac output (CO) and other hemodynamic parameters in patients undergoing gynecological laparoscopic surgery in head-down lithotomy position and Trendelenburg position. Methods Sixty patients were divided into head-down lithotomy group and Trendelenburg group. CO was recorded as baseline by a noninvasive cardiac output monitor NICOM? system after the placement of patients. These measurements were also acquired when the patients were placed in the 30° head-down tilt(T0)following pneumoperitoneum establishment.Stroke volume(SV), heart rate(HR)and CO were monitored at 1-minute intervals thereafter for a total of 10 minutes(T1-T10),and mean arterial pressure(MAP)and total peripheral resistance(TPR)were monitored every 5 minutes. Results The reduction of CO in head-down lithotomy group was greater than that in Trendelenburg group(T0:-31％±19％ vs.-9％±34％;T1:-32％±18％ vs.-16％±38％;T2:-33％± 19％vs.-16％±26％;T3:-32％±22％vs.-16％±28％;T4:-31％±18％vs.-12％±38％;T5:-30％± 17％vs.-14％±37％;T6:-31％±17％ vs.-14％±33％,all P<0.05)during the first 6 minutes. MAP at baseline in head-down lithotomy group was significantly higher than that in Trendelenburg group[(97±11) mmHg vs.(85±6)mmHg,P<0.05].MAP decreased in head-down lithotomy group at T0(-8％±16％) and increased in Trendelenburg group at T5 and T10(T5:9％±15％,T10:12％±18％). Conclusion CO reduction was greater in patients in head-down lithotomy position than that in Trendelenburg position group during the first 10 minutes after adjusting the position following pneumoperitoneum establishment.
ABSTRACT
Objective:To investigate the proliferation and apoptosis characteristics of polyploid non-small cell lung cancer (NSCLC) cell model induced by docetaxel (Doc), and to analyze the potential role of polyploid tumor cells in chemotherapy resistance and tumor recurrence.Methods:NSCLC A549 cells were treated with dimethyl sulfoxide (DMSO) or 1 μmol/L Doc for 24 h. After drug removal, the cells were cultured in complete medium until the third day or the 5th day, and then they were recorded as the control group, Doc 24 h group, Doc 24 h+ 3 d group, Doc 24 h + 5 d group, respectively. The cell morphology was detected by using immunofluorescence staining. Flow cytometry was used to determine cell ploidy and cell cycle. Dil labeling and CFSE labeling were applied to detect cell proliferation. Flow cytometry by Annexin-V/PI double labeling was used to detect apoptosis. The changes of cyclin and apoptotic protein were analyzed by using Western blot.Results:Immunofluorescence staining results showed that compared with the control group, the volume of a small number of surviving cells in Doc 24 h group was increased slightly and the cells showed multinuclear status; while the cell volume in Doc 24 h+ 3 d group and Doc 24 h+ 5 d group continued to increase, and the nucleus remained multinuclear. The results of cell ploidy analysis also showed that the percentage of polyploid cell subsets was (3.40±0.95)%, (20.80±2.87)% in Doc 24 h group, (55.67±3.85)% in Doc 24 h+3 d group and (76.20±2.51)% in Doc 24 h+5 d group. With the prolongation of withdrawal time, the percentage of polyploid cell subsets was increased, and the difference was statistically significant ( F= 478.054, P < 0.05). The percentage of G 1 and S phase cell subsets in Doc 24 h group was lower than that in the control group, and the percentage of G 2/M phase cell subsets was higher than that in the control group, and the difference was statistically significant (both P < 0.05). The protein expression level of cdc2, P-cdc2 (Thr14), P-cdc2 (Tyr15), P-cyclin B1 (Ser128), P-cyclin B1 (Ser147) in the cells of the control group, Doc 24 h group, Doc 24 h+ 3 d group and Doc 24 h+ 5 d group was down-regulated in sequence, while the expression level of cyclin B1 was up-regulated, and cdc25c was down-regulated in Doc 24 h + 3 d group and Doc 24 h+ 5 d group. Dil staining results showed that the fluorescence of cell-labeled Dil in Doc 24 h group, Doc 24 h+ 3 d group and Doc 24 h + 5 d group did not decrease significantly. CFSE staining showed that the fluorescence intensity of CFSE labeled by polyploid A549 cells did not change significantly with the prolonged withdrawal time. Annexin-V/PI double staining showed that the percentage of apoptotic cell subsets in Doc 24 h group was higher than that in the control group ( P < 0.05), but the percentage of apoptotic cell subsets in Doc 24 h + 3 d group and Doc 24 h + 5 d group was lower than that in Doc 24 h group, while there was no statistically significant difference when compared with the control group ( P > 0.05). Western blot results showed that the expression of bcl-xl and mcl-1 in the control group, Doc 24 h group, Doc 24 h + 3 d group and Doc 24 h + 5 d group was up-regulated in sequence, while the expression of bax and bak in Doc 24 h + 3 d group and Doc 24 h + 5 d group was up-regulated, but down-regulated in Doc 24 h+5 d group. Conclusions:Doc can induce polyploidy of A549 cells in vitro. The cell cycle is blocked in G 2/M phase. After doc treatment, the proliferation of A549 cells is significantly decreased, and the apoptosis of A549 cells is promoted. However, with the prolongation of withdrawal time, apoptosis resistance occurs, and the expression levels of corresponding pro-apoptosis and anti-apoptosis proteins show significant changes. This may be helpful for polyploid tumor cells to produce drug resistance and tumor recurrence after chemotherapy intervention.
ABSTRACT
Objective:To explore the influencing factors of rapid postoperative recovery in young(≤40 years old) lung cancer patients.Methods:Retrospective analysis was performed on 82 young patients with lung cancer diagnosed by postoperative pathology admitted to the department of thoracic surgery of the first affiliated hospital of Zhengzhou University from March 2013 to March 2019, the patients were divided into two groups according to their postoperative hospitalization time(hospitalization time≤7d, hospitalization time >7d). The preoperative medical history and examination data, intraoperative(operative method, embedding materials), postoperative complications and postoperative treatment and other data of the enrolled patients were collected to analyze the relationship between various factors and postoperative hospitalization time.Univariate analysis used t test or Fisher exact probability method, multivariate analysis used logistic regression model to analyze the data . Results:All 82 patients successfully completed the operation, and no death occurred during the perioperative period. There were no significant differences( P>0.05)according to the two groups of patients in the preoperative pulmonary function(FEV1) operation history, history of hypertension, diabetes, history of preoperative chemotherapy and surgery in the patients' position, blood transfusion, pleural adhesion, Czech, nai d, the use of xanthan gum, operation time, the maximum diameter and postoperative tumor thermal perfusion, fever, vomiting, choking cough, abdominal distension, etc.And it has significant differences( P<0.05). In the preoperative antibiotic use( P=0.002), the improvement of lung function( P=0.018), smoking history( P=0.024), medical reasons( P=0.011) and the operation( P<0.001), the lymph node excision scope( P<0.001), the lymph node dissection( P=0.017), hemostatic material use( P=0.023), blood loss( P=0.001) and postoperative average white blood cell count( P=0.033). Conclusion:Preoperative and postoperative prophylactic use of antibiotics and drugs to improve pulmonary function were beneficial to postoperative recovery.Smoking was an independent risk factor for prolonged postoperative hospital stay.Minimally invasive operation and application of hemostatic materials can effectively shorten the postoperative hospitalization time of patients.
ABSTRACT
Objective:To investigate the expression of coxsackievirus-adenovirus receptor (CAR) in thymic carcinoma and the relationship between CAR and the antitumor activity of oncolytic adenovirus H101.Methods:The expression of CAR in thymic carcinoma tissues and cells were detected by RT-qPCR and Western blot. H101 expression and virus titers in Bcap-37, MP59 and T1889 cells after infection were detected by RT-qPCR and 50% tissue culture infectious dose (TCID 50). The proliferation activity and apoptosis rates of T1889 cells infected with H101 at different multiplicity of infection (MOI) were detected by CCK-8 and flow cytometry. CAR expression in T1889 cells treated with different concentrations of trichostatin A (TSA), a histone deacetylase inhibitor, was detected. H101 expression and virus titers in the TSA-treated and H101-infected cells were detected. Cell activity was detected by CCK-8. The phosphorylation levels of MARK and ERK1/2 and the expression of CAR at protein level in TSA-treated or TSA+ TBHQ (ERK activator) treated cells were detected. Results:CAR expression at both mRNA and protein levels were significantly lower in thymic carcinoma tissues than in adjacent normal tissues ( P<0.01), and lower in MP59 and T1889 cells than in thymic epithelial cells (TEC) and Bcap-37 cells ( P<0.01). H101 expression in MP59 and T1889 cells and the titers of H101 in culture supernatants were significantly lower than those in Bcap-37 cells ( P<0.01). Compared with Bcap-37 cell, the activity of MP59 and T1889 cells was significantly increased and the apoptosis rates were significantly decreased 48 h after H101 infection ( P<0.01). The expression of CAR at both mRNA and protein levels in T1889 cells treated with different concentrations of TSA increased in a dose-dependent manner. When T1889 cells were treated with 0.25 μmol/L of TSA, the expression of H101 at mRNA level and H101 titers were significantly increased ( P<0.05); the phosphorylation levels of MAPK and ERK1/2 proteins were continuously decreased; the expression of CAR was continuously increased. Compared with the TSA treatment group, the expression of CAR at protein level in the TSA+ TBHQ treatment group decreased significantly ( P<0.01), and the p-ERK1/2/ERK1/2 ratio increased significantly ( P<0.01). Conclusions:TSA could up-regulate CAR expression in thymic carcinoma by inhibiting the MARK/ERK1/2 pathway, thereby enhancing the antitumor activity of H101.
ABSTRACT
Objective@#To explore the influencing factors of rapid postoperative recovery in young(≤40 years old) lung cancer patients.@*Methods@#Retrospective analysis was performed on 82 young patients with lung cancer diagnosed by postoperative pathology admitted to the department of thoracic surgery of the first affiliated hospital of Zhengzhou University from March 2013 to March 2019, the patients were divided into two groups according to their postoperative hospitalization time(hospitalization time≤7d, hospitalization time >7d). The preoperative medical history and examination data, intraoperative(operative method, embedding materials), postoperative complications and postoperative treatment and other data of the enrolled patients were collected to analyze the relationship between various factors and postoperative hospitalization time.Univariate analysis used t test or Fisher exact probability method, multivariate analysis used logistic regression model to analyze the data .@*Results@#All 82 patients successfully completed the operation, and no death occurred during the perioperative period. There were no significant differences(P>0.05)according to the two groups of patients in the preoperative pulmonary function(FEV1) operation history, history of hypertension, diabetes, history of preoperative chemotherapy and surgery in the patients' position, blood transfusion, pleural adhesion, Czech, nai d, the use of xanthan gum, operation time, the maximum diameter and postoperative tumor thermal perfusion, fever, vomiting, choking cough, abdominal distension, etc.And it has significant differences(P<0.05). In the preoperative antibiotic use(P=0.002), the improvement of lung function(P=0.018), smoking history(P=0.024), medical reasons(P=0.011) and the operation(P<0.001), the lymph node excision scope(P<0.001), the lymph node dissection(P=0.017), hemostatic material use(P=0.023), blood loss(P=0.001) and postoperative average white blood cell count(P=0.033).@*Conclusion@#Preoperative and postoperative prophylactic use of antibiotics and drugs to improve pulmonary function were beneficial to postoperative recovery.Smoking was an independent risk factor for prolonged postoperative hospital stay.Minimally invasive operation and application of hemostatic materials can effectively shorten the postoperative hospitalization time of patients.
ABSTRACT
Objective To investigate the spatio-temporal distribution characteristics of Oncomelania hupensis snail habitats in three cities of Suzhou, Wuxi and Changzhou along the Taihu Lake region, so as to provide technical supports for establishing a sensitive and highly effective surveillance and forecast system for schistosomiasis. Methods Snail distribution data were collected from Suzhou, Wuxi and Changzhou cities from 1950 to 2018, and the changing trend for snail habitats were described over years. In addition, the clusters of snail habitats were detected using Kernel density analysis and SaTScan space-time scan analysis. Results The number of snail habitats appeared a single-peak distribution in Suzhou, Wuxi and Changzhou cities from 1950 to 2018, which peaked in 1970 and then declined rapidly. There were 62.68% of snail habitats eliminated within 10 years after identification, of which 38.24% were eliminated at the year of identification. Kernel density analysis and SaTScan space-time scan analysis revealed that high-density clusters of snail habitats were mainly distributed in Kunshan City, Wuzhong District and Xiangcheng District from 1970 to 1980, and in Yixing City in 1990; since then, the clusters gradually shrank, and overall appeared a move from northeast to west of Taihu Lake. A total of 4 new clusters were detected after 1970, as revealed by space-time scanning of snail habitats. In current snail habitats, emerging snail habitats are mainly identified in Huqiu District (Dongzhu Town), Wuzhong District (Guangfu Town), Taicang City (Shaxi Town) and Jintan District, and re-emerging snail habitats are scattered in 7 districts. Conclusions The distribution of snail habitats are spatio-temporal aggregation in Suzhou, Wuxi and Changzhou cities. The monitoring and prediction of emerging and re-emerging snail habitats are the key points in the future.
ABSTRACT
Objective To establish a recombinase-aided isothermal amplification (RAA) assay for nucleic acid detection of Schistosoma mansoni. Methods The 121 bp highly-repeated sequence of S. mansoni was selected as the target gene fragment to be detected. The primers and fluorescent probes were designed using the Amplfix software, and a fluorescent RAA assay was established and optimized. The fluorescent RAA assay was performed to detect gradient diluent recombinant plasmids containing target gene fragment and different concentrations of S. mansoni genomic DNA to determine the sensitivity, and this assay was applied to detect the genomic DNA of S. japonicum, S. haematobium, Ancylostoma duodenale and Clonorchis sinensis to evaluate the specificity. Results A fluorescent RAA assay was successfully established, which was effective to amplify the specific gene fragments of S. mansoni within 20 min at 39 ℃. The minimum detectable limit of the fluorescent RAA assay was 10 copies/μL using recombinant plasmids as templates and 0.1 fg/μL using S. mansoni genomic DNA samples as templates. The fluorescent RAA assays were all negative for detecting the genomic DNA from S. japonicum, S. haematobium, A. duodenale and C. sinensis. Conclusions A novel fluorescent RAA assay is successfully established, which is simple, rapid, sensitive and specific to detect genomic DNA of S. mansoni.
ABSTRACT
Objective@#To investigate the effects of human umbilical cord mesenchymal stem cells (MSCs) on the proliferation, apoptosis, migration, invasion and stemness of esophageal cancer EC1 cells.@*Methods@#Human umbilical cord mesenchymal stem cells were isolated and cultured in vitro, and cell phenotype was identified by flow cytometry. MSCs or their conditioned medium were co-cultured with esophageal cancer EC1 cells. The effects on the proliferation, apoptosis, migration, invasion and stemness of EC1 cells were examined by cell counting kit-8 (CCK-8), flow cytometry, Transwell, quantitative real-time polymerase chain reaction (RT-qPCR) and spheroid formation assays.@*Results@#MSCs inhibited the proliferation of EC1 cells in a concentration dependent manner. When the ratio of MSCs to EC1 cells was 0∶1, 1∶1, 2∶1, 5∶1, the apoptotic rates of EC1 cells were (4.07±0.34)%, (8.90±0.36)%, (10.80±0.50)% and (15.23±1.06)%, respectively, suggesting that MSCs promoted the apoptosis of EC1 cells in a concentration dependent manner (all P<0.05). The expression levels of OCT2, SOX2, KLF4, CXCR4 and CXCR7 in EC1 cells cultured in 80% conditioned medium were 0.53±0.03, 0.49±0.02, 0.73±0.09, 0.57±0.05 and 0.24±0.02, respectively, which were lower than those in the regular medium group (all P<0.05). The numbers of migrated cells in regular medium as well as 10%, 40%, and 80% conditioned medium were 287.3±21.6, 280.7±15.5, 264.3±16.8, and 257.7±8.0, respectively. Meanwhile, the numbers of invasive cells were 194.3±16.6, 213.7±24.3, 221.0±16.0, (252.0±20.4), respectively. There was no significant difference between the groups (all P>0.05).@*Conclusion@#Human umbilical cord mesenchymal stem cells can inhibit the proliferation, promote apoptosis and reduce the stemness, and have no significant effect on the migration and invasion of EC1 cells.