Study on changes of proliferation, migration and invasion abilities of radiation-induced polyploid colon cancer SW1116 cells and their progeny and the related mechanisms / 肿瘤研究与临床

Objective:To investigate the characteristics related to proliferation, migration and invasion of radiation-induced polyploid colon cancer SW1116 cells and their progeny.Methods:Colon cancer SW1116 cells were conventionally cultured in Leibovitz's L-15 medium containing 10% fetal bovine serum. SW1116 cells at logarithmic growth stage were irradiated with 7 Gy X-ray, and the morphological changes of the cells were observed by inverted microscope on days 3, 5, 10 and 19 after radiation induction. According to the morphological changes of the cells, the cells at day 3 after radiation induction were labeled as polyploid giant cancer cell (PGCC) group, and the cells at day 19 were recorded as PGCC progeny group. SW1116 cells without radiation induction were used as control group. Flow cytometry was used to detect cell ploidy in the control, PGCC and PGCC progeny groups, CCK-8 assay was used to detect the proliferation ability of the three groups, cell migration and invasion abilities of the three groups were detected by cell scratch assay and Transwell assay, and Western blotting was used to detect the expressions of cell cycle and proliferation-related proteins and epithelial-mesenchymal transition (EMT) marker N-cadherin (N-cad) in the three groups.Results:The volume of SW1116 cells gradually became larger on days 3, 5 and 10 after radiation induction, and returned to normal on day 19. The proportions of polyploid (DNA content >4N) cell subsets in the control group, PGCC group and PGCC progeny group were (2.3±1.1)%, (23.1±8.1)% and (3.2±0.5)%, the difference was statistically significant ( F = 18.52, P < 0.05), and the proportion of polyploid cell subpopulations in the PGCC group was higher than that in the control group ( t = 5.38, P < 0.01), but the differences between the PGCC progeny group and the control group were not statistically significant ( t = 0.22, P > 0.05). After 72 h of culture, the cell proliferation rates of the control, PGCC and PGCC progeny groups were (100.0±4.1)%, (73.5±0.7)% and (123.9±3.5)%, and the difference was statistically significant ( F = 190.27, P < 0.001). After 48 h of cell scratching, the scratch healing rates in the control, PGCC and PGCC progeny groups were (38.0±2.7)%, (41.5±4.0)% and (63.7±4.2)%, and the difference was statistically significant ( F = 43.05, P < 0.001). After 24 h of culture, the number of invasive cells in the control, PGCC and PGCC progeny groups was 12.9±1.2, 3.4±0.6 and 23.7±1.5, and the difference was statistically significant ( F = 63.64, P < 0.001). The expression levels of cell cycle-related proteins P-cdc25c, cdc25c and cdc2 in the PGCC group were lower than those in the control group (all P < 0.05), and the expression levels of transcription factor-related proteins E2F-2, E2F-3 and EMT marker N-cad were downregulated compared with the control group (all P < 0.05); the expression levels of P-cdc25c, cdc25c, cdc2, E2F-2, E2F-3 and N-cad proteins in the PGCC progeny group were higher than those in the control group (all P < 0.05). Conclusions:Radiation can induce colon cancer SW1116 cells to produce polyploid, which may then generate daughter cells through asymmetric mitosis and gain new life, and then promote the recurrence and metastasis of colon cancer.

An Optogenetics-based Approach to Regulate Colonic Contractions by Modulating the Activity of the Interstitial Cells of Cajal in Mice

Background/Aims@#The interstitial cells of Cajal (ICC) are pacemaker cells in the gastrointestinal (GI) tract. We examined whether the activity of ICC could be stimulated to control colonic contractions. An optogenetics-based mouse model in which the light-sensitive protein channelrhodopsin-2 (ChR2) was expressed was used to accomplish cell specific, direct stimulation of ICC. @*Methods@#An inducible site-specific Cre-loxP recombination system was used to generate KitCreERT2/+ ;ROSAChR2(H134R)/tdTomato/+ mice in which ChR2(H134R), a variant of ChR2, was genetically expressed in ICC after tamoxifen administration. Genotyping and immunofluorescence analysis were performed to confirm gene fusion and expression. Isometric force recordings were performed to measure changes in contractions in the colonic muscle strips. @*Results@#ChR2 was specifically expressed in Kit-labeled ICC. The isometric force recordings showed that the contractions of the colonic muscle strips changed under 470 nm blue light. Light stimulation evoked premature low-frequency and high amplitude (LFHA) contractions and enhanced the frequency of the LFHA contractions. The light-evoked contractions were blocked by T16Ainh-A01, an antagonist of anoctamin 1 channels that are expressed selectively in ICC in colonic muscles. @*Conclusions@#Our study demonstrates a potentially feasible approach to stimulate the activity of ICC by optogenetics. The colonic motor patterns of muscle strips, especially LFHA contractions, can be regulated by 470 nm light via ChR2, which is expressed in ICC.

Establishment and evaluation of a method for extracting exogenous short DNA fragments of Schistosoma japonicum from urine samples / 中国血吸虫病防治杂志

Objective To establish the method for extracting exogenous short DNA fragments of Schistosoma japonicum from urine samples, and to evaluate the efficiency of this method for extraction from urine samples treated with various methods. Methods The S. japonicum SjG28 gene fragment was selected as a target sequence, and the 81 bp short DNA fragment was amplified on the target sequence using PCR assay. Following characterization using sequencing, the short DNA fragment was added into the urine samples as an exogenous short DNA fragment. Primers and probes were designed with SjG28 as a target gene, to establish the real-time fluorescent quantitative PCR (qPCR) assay. The sensitivity of this qPCR assay was evaluated with exogenous short DNA fragments that were diluted at a 1:10 dilution ratio as the DNA template, and the specificity of the qPCR assay was evaluated with the genomic DNA of S. mansoni, S. haematobium, Babesia, Ancyiostoma duodenaie, Cionorchis sinensis, and Paragonimus westermani as DNA templates. Exogenous short DNA fragments were added into artificial and healthy volunteers’ urine samples, followed by pH adjustment, centrifugation and concentration, and the efficiency of extracting exogenous short DNA fragments from urine samples was compared with the QIAmp Viral RNA Mini Kit (Qiagen kit) and BIOG cfDNA easy kit (BIOG kit). Results An 81 bp small DNA fragment of S. japonicum was successfully prepared, and the lowest detection limit of the established qPCR assay was 100 copies/μL of the 81 bp small DNA fragment of S. japonicum. If the genomic DNA of S. japonicum, S. mansoni, S. haematobium, Babesia, A. duodenaie, C. sinensis, and P. westermani served as DNA templates, the qPCR assay only detected fluorescent signals with S. japonicum genomic DNA as the DNA template. If the pH values of artificial urine samples were adjusted to 5, 6, 7 and 8, the recovery rates were (49.12 ± 2.09)%, (84.52 ± 4.96)%, (89.38 ± 3.32)% and (87.82 ± 3.90)% for extracting the exogenous short DNA fragment of S. japonicum with the Qiagen kit, and were (2.30 ± 0.07)%, (8.11% ± 0.26)%, (13.35 ± 0.61)% and (20.82 ± 0.68)% with the BIOG kit, respectively (t = 38.702, 26.955, 39.042 and 29.571; all P values < 0.01). If the Qiagen kit was used for extracting the exogenous short DNA fragment from artificial urine samples, the lowest recovery rate was seen from urine samples with a pH value of 5 (all P values < 0.05), and there were no significant differences in the recovery rate from urine samples with pH values of 6, 7 and 8 (all P values > 0.05). Following centrifugation of artificial [(64.30 ± 1.00)% vs. (58.87 ± 0.26)%; t = 12.033, P < 0.05] and healthy volunteers’ urine samples [(31 165 ± 1 017) copies/μL vs. (28 471 ± 818) copies/μL; t = 23.164, P < 0.05]. In addition, concentration of artificial urine samples with the 10 kDa Centrifugal Filter and concentration of healthy volunteers’ urine samples with the 100 kDa Centrifugal Filter were both effective to increase the recovery of the Qiagen kit for extracting the exogenous short DNA fragment of S. japonicum (both P values < 0.01). Conclusions A method for extracting exogenous short DNA fragments of S. japonicum from urine samples has been successfully established, and the Qiagen kit has a high extraction efficiency. Adjustment of urine pH to 6 to 8 and concentration of healthy volunteers’ urine samples with the 100 kDa Centrifugal Filter are both effective to increase the efficiency of extracting exogenous short DNA fragments of S. japonicum.

Protective effect of melatonin against oxygen-induced retinopathy: a study based on the HMGB1/NF-κB/NLRP3 axis / 中国当代儿科杂志

OBJECTIVES@#To study the protective effect of melatonin (Mel) against oxygen-induced retinopathy (OIR) in neonatal mice and the role of the HMGB1/NF-κB/NLRP3 axis.@*METHODS@#Neonatal C57BL/6J mice, aged 7 days, were randomly divided into a control group, a model group (OIR group), and a Mel treatment group (OIR+Mel group), with 9 mice in each group. The hyperoxia induction method was used to establish a model of OIR. Hematoxylin and eosin staining and retinal flat-mount preparation were used to observe retinal structure and neovascularization. Immunofluorescent staining was used to measure the expression of proteins and inflammatory factors associated with the HMGB1/NF-κB/NLRP3 axis and lymphocyte antigen 6G. Colorimetry was used to measure the activity of myeloperoxidase.@*RESULTS@#The OIR group had destruction of retinal structure with a large perfusion-free area and neovascularization, while the OIR+Mel group had improvement in destruction of retinal structure with reductions in neovascularization and perfusion-free area. Compared with the control group, the OIR group had significant increases in the expression of proteins and inflammatory factors associated with the HMGB1/NF-κB/NLRP3 axis, the expression of lymphocyte antigen 6G, and the activity of myeloperoxidase (P<0.05). Compared with the OIR group, the OIR+Mel group had significant reductions in the above indices (P<0.05). Compared with the control group, the OIR group had significant reductions in the expression of melatonin receptors in the retina (P<0.05). Compared with the OIR group, the OIR+Mel group had significant increases in the expression of melatonin receptors (P<0.05).@*CONCLUSIONS@#Mel can alleviate OIR-induced retinal damage in neonatal mice by inhibiting the HMGB1/NF-κB/NLRP3 axis and may exert an effect through the melatonin receptor pathway.

Enzalutamide and olaparib synergistically suppress castration-resistant prostate cancer progression by promoting apoptosis through inhibiting nonhomologous end joining pathway / 亚洲男科学杂志(英文版)

Recent studies revealed the relationship among homologous recombination repair (HRR), androgen receptor (AR), and poly(adenosine diphosphate-ribose) polymerase (PARP); however, the synergy between anti-androgen enzalutamide (ENZ) and PARP inhibitor olaparib (OLA) remains unclear. Here, we showed that the synergistic effect of ENZ and OLA significantly reduced proliferation and induced apoptosis in AR-positive prostate cancer cell lines. Next-generation sequencing followed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed the significant effects of ENZ plus OLA on nonhomologous end joining (NHEJ) and apoptosis pathways. ENZ combined with OLA synergistically inhibited the NHEJ pathway by repressing DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and X-ray repair cross complementing 4 (XRCC4). Moreover, our data showed that ENZ could enhance the response of prostate cancer cells to the combination therapy by reversing the anti-apoptotic effect of OLA through the downregulation of anti-apoptotic gene insulin-like growth factor 1 receptor ( IGF1R ) and the upregulation of pro-apoptotic gene death-associated protein kinase 1 ( DAPK1 ). Collectively, our results suggested that ENZ combined with OLA can promote prostate cancer cell apoptosis by multiple pathways other than inducing HRR defects, providing evidence for the combined use of ENZ and OLA in prostate cancer regardless of HRR gene mutation status.

Clinical evidence-based guideline for the diagnosis and treatment of anterior cruciate ligament injury (2022 version) / 中华创伤杂志

The anterior cruciate ligament (ACL) injury is a common sports injury that has a significant impact on knee function and patients′ mobility. With the popularity of national fitness campaign in China, the incidence of ACL injury is increasing year by year. Currently, there still lacks clinical standards or guidelines on how to choose appropriate treatment methods, surgical plans and rehabilitation protocols for ACL injury. In order to timely reflect the new treatment concept of ACL injury, standardize its diagnosis and treatment and improve the curative effect, the Sports Medicine Society of Chinese Research Hospital Association and the Editorial Board of Chinese Journal of Trauma organized domestic orthopedic and sports medicine experts to formulate the "clinical evidence-based guideline for the diagnosis and treatment of anterior cruciate ligament injury (2022 version)" based on the level of evidence-based medicine and in compliance with the principle of scientificity, practicability and advancement. The present guideline includes 12 recommendations for the diagnosis, treatment and rehabilitation of ACL injury in order to provide guidance and assistance for the clinical diagnosis and treatment of ACL injury in China.

Effects of arsenic, selenium, lead and chromium in urine on blood pressure in people from arsenic exposure areas of Togtoh County, Inner Mongolia Autonomous Region / 中华地方病学杂志

Objective:To learn about the incidence of hypertension in residents of arsenic exposure areas of Togtoh County, Inner Mongolia Autonomous Region, and to analyze the influencing factors of hypertension.Methods:In Inner Mongolia Autonomous Region, Ciweigou of Togtoh County, a drinking water-type endemic arsenic poisoning area, and Lanjiayao of Horinger County, a non-arsenic poisoning area with similar living habits and economic conditions, permanent residents who had lived for ≥10 years were selected as the survey subjects. Totally 116 residents of Ciweigou (exposure group) and 68 residents of Lanjiayao (control group) were included in the survey. Blood pressure was measured and the contents of arsenic, selenium, lead and chromium in urine were detected, multivariate logistic regression was used to analyze the influencing factors of hypertension.Results:The detection rates of hypertension in exposure group and control group were 53.45% (62/116) and 70.59% (48/68), respectively, and the difference between the two groups was statistically significant (χ 2 = 5.33, P = 0.022). The contents of arsenic, selenium and chromium in urine of exposure group were higher than those of control group, and the content of lead in urine was lower than that of control group, and the differences were statistically significant ( Z = 13.04, 6.34, 11.28, - 9.91, P < 0.001). The results of multivariate logistic regression analysis showed that female, age ≥60 years old and high urinary arsenic content were the influencing factors of hypertension [odds ratio ( OR) = 2.074, 2.004, 0.424, 95% confidence interval ( CI): 1.113 - 3.866, 1.035 - 3.879, 0.219 - 0.820] in arsenic exposure areas. Conclusion:Female, age ≥60 years old and high urinary arsenic content are the influencing factors of hypertension in arsenic exposure areas of Togtoh County, Inner Mongolia Autonomous Region.

Progress in the diagnosis of minimal hepatic encephalopathy / 中华肝脏病杂志

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.

Effects of human umbilical cord-derived mesenchymal stem cells and their conditioned medium on proliferation, migration and apoptosis of non-small cell lung cancer polyploid A549 cells / 肿瘤研究与临床

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.

A Fluorescent Recombinase Aided Amplification Assay for Detection of Babesia microti

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.

Chinese thoracic surgery experts consensus on postoperative follow-up plans for esophageal squamous cell carcinoma / 中国胸心血管外科临床杂志

@#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.

Development of a fluorescent recombinase-aided isothermal amplification-based nucleic acid assay for detection of Leishmania / 中国血吸虫病防治杂志

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.

Establishment of a nucleic acid dipstick test for detection of Schistosoma japonicum specific gene fragments based on the recombinase-aided isothermal amplification assay / 中国血吸虫病防治杂志

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.

Efficacy analysis of Da Vinci robotic assisted and laparoscopic assisted complete mesocolic excision for right hemicolon cancer / 中华消化外科杂志

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.

Discovery of 4-arylthiophene-3-carboxylic acid as inhibitor of ANO1 and its effect as analgesic agent

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

Performance of a recombinase - aided amplification assay for detection of Schistosoma japonicum infections in Oncomelania hupensis / 中国血吸虫病防治杂志

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.

Study on the proliferation and apoptosis characteristics of polyploid non-small cell lung cancer A549 cells induced by docetaxel / 肿瘤研究与临床

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.

Analysis of influencing factors for rapid postoperative recovery of young lung cancer patients / 中华胸心血管外科杂志

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.

Trichostatin A up-regulates coxsackievirus-adenovirus receptor (CAR) expression through inhibiting MAPK/ERK pathway to enhance the antitumor activity of H101 virus in thymic carcinoma / 中华微生物学和免疫学杂志

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.

Construction of prediction model of celiac lymph node metastasis in thoracic esophageal squamous cell carcinoma and risk subgroup analysis of celiac lymph node metastasis probability / 中华消化外科杂志

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.