BACKGROUND:In the research and application of neural stem cells,cell culture and passage are key links,which directly affect the quality of cells and experimental results.It is of great significance to find the most suitable digestive enzymes that can maintain the biological characteristics of embryonic mouse spinal cord neural stem cells and enhance their passage efficiency.OBJECTIVE:To explore the most suitable digestive enzyme for passage of neural stem cells from the spinal cord of embryonic mice.METHODS:Microscopic dissection was used to isolate and extract spinal cord tissue from E14 d embryonic mice,which was cultured in DMEM/F12 serum-free medium containing epidermal growth factor,basic fibroblast growth factor,and B27.After spherulation,Nestin and Sox2 immunofluorescence identification was performed.During neural stem cell passage and culture,single-cell suspensions were prepared using trypsin,papain,and TrypLETM Express enzyme digestion combined with blow molding.The cell dispersion and spheroidization were observed,and passage 3 cells were stained with propidium iodide to detect cell death.Cell proliferation was detected by counting the total number of cells.Immunofluorescence staining,western blot assay and RT-PCR were used to detect the expression of Olig2,Tuj1,GFAP,and NeuN at the protein and mRNA levels and to identify cell differentiation.RESULTS AND CONCLUSION:After 72 hours of culture,E14 d embryonic mouse spinal cord tissue cells could form suspended neurospheres,which could be passaged after 5-7 days.Compared with trypsin and papain,TrypLETM Express enzyme combined with blow beating method was used for passage.The cell dispersion rate was high,the activity was good,and more NeuN-and Tuj1-positive neurons differentiated.This study optimized the culture and passaging process of neural stem cells,laying a foundation for further research on stem cell transplantation therapy for spinal cord diseases.

BACKGROUND:In the research and application of neural stem cells,cell culture and passage are key links,which directly affect the quality of cells and experimental results.It is of great significance to find the most suitable digestive enzymes that can maintain the biological characteristics of embryonic mouse spinal cord neural stem cells and enhance their passage efficiency.OBJECTIVE:To explore the most suitable digestive enzyme for passage of neural stem cells from the spinal cord of embryonic mice.METHODS:Microscopic dissection was used to isolate and extract spinal cord tissue from E14 d embryonic mice,which was cultured in DMEM/F12 serum-free medium containing epidermal growth factor,basic fibroblast growth factor,and B27.After spherulation,Nestin and Sox2 immunofluorescence identification was performed.During neural stem cell passage and culture,single-cell suspensions were prepared using trypsin,papain,and TrypLETM Express enzyme digestion combined with blow molding.The cell dispersion and spheroidization were observed,and passage 3 cells were stained with propidium iodide to detect cell death.Cell proliferation was detected by counting the total number of cells.Immunofluorescence staining,western blot assay and RT-PCR were used to detect the expression of Olig2,Tuj1,GFAP,and NeuN at the protein and mRNA levels and to identify cell differentiation.RESULTS AND CONCLUSION:After 72 hours of culture,E14 d embryonic mouse spinal cord tissue cells could form suspended neurospheres,which could be passaged after 5-7 days.Compared with trypsin and papain,TrypLETM Express enzyme combined with blow beating method was used for passage.The cell dispersion rate was high,the activity was good,and more NeuN-and Tuj1-positive neurons differentiated.This study optimized the culture and passaging process of neural stem cells,laying a foundation for further research on stem cell transplantation therapy for spinal cord diseases.

Objective: To explore the valuable indicators for differential diagnosis by comparing the clinical features of Crohn′s disease (CD) with primary intestinal lymphoma (PIL). Methods: From 2010 to 2017, at The Seventh Medical Center of PLA General Hospital, a total of 91 patients diagnosed with CD or PIL were enrolled, including 76 cases of CD, 14 cases of PIL and one case of CD with secondary lymphoma. The clinical data of enrolled patients were retrospectively analyzed. T-test, non-parametric test and chi-square test were used for statistical analysis. Results: The average age at diagnosis of CD patients was (37.7±16.0) years, which was younger than that of PIL patients (52.6±19.6) years, and the difference was statistically significant (t=-3.085, P=0.003). The median duration of CD was 36.0 months, which was longer than that of PIL (3.5 months), and the difference was statistically significant (Z=-3.616, P<0.01). Abdominal pain and extra-intestinal manifestations (oral ulcers, joint injuries, erythema nodosum and perianal lesions) were more common in CD patients, and the differences were all statistically significant (χ²=9.427 and 5.173, both P<0.05). CD patients were not diagnosed by colonoscopic biopsy alone. Totally 13 of 14 patients were diagnosed by colonoscopic biopsy and immunohistochemisty and one patient was diagnosed after surgery. The diagnostic rate of pathological biopsy in PIL group was significantly higher than that in CD group (χ²=82.584, P<0.01). One patient was initially clinically diagnosed as CD, and then developed secondary lymphoma after five times of infliximab treatment. Conclusions The diagnosis of CD is generally supported by young age, long course of disease, abdominal pain and extra-intestinal manifestations. The diagnostic rate of colonoscopic biospsy in PIL patients is higher than that in CD patients.

BACKGROUND: Patients with lung cancer are often accompanied by anxiety, which affects postoperative recovery. The aim of this study is to explore the effects of preoperative anxiety on early prognosis in patients after thoracoscopic lung cancer resection. METHODS: A total of 100 patients undergoing thoracoscopic resection of lung cancer were divided into 2 groups by hospital anxiety and depression scale (HADS): 44 in anxiety group (anxiety score>8) and 56 in control group (anxiety score<8). The primary endpoint: length of postoperative hospital stay. The secondary endpoint: length of hospital stay, visual analogue scale (VAS), the incidence of nausea and vomiting as well as postoperative new arrhythmia and the consumption of postoperative analgesic and rescue antiemetic. RESULTS: Compared with the control group, the length of postoperative hospital stay and hospital stay in the anxiety group were both significantly longer [(5.1±2.5) d vs (4.0±1.3) d, P<0.01; (10.9±4.0) d vs (9.1±4.1) d, P<0.05)], the VAS score and the incidence of nausea as well as arrhythmia were significantly increased [(4.7±1.9) vs (2.6±1.8), P<0.001; 40.9% vs 16.1%, P<0.01; 36.4% vs 20.7%, P<0.05], and the consumption of postoperative analgesic and rescue antiemetic were also significantly increased [(72.5±8.9) mL vs (68.2±9.4) mL, P<0.05; (2.1±2.9) mg vs (0.9±1.9) mg, P<0.05]. CONCLUSIONS Preoperative anxiety can affect the early prognosis of patients after thoracoscopic lung cancer resection, prolong hospitalization time, increase the postoperative pain score and the incidence of postoperative nausea and new arrhythmia as well as the consumption of postoperative analgesic and rescue antiemetic.

While the near infrared spectroscopy (NIRS) is used to measure the inhomogeneous samples with diffuse reflection mode, the accuracy and robustness of the calibration model is not quite good for the variation of spectrum scattering and absorption coefficient in those samples.Therefore, an establishment strategy of hybrid model based on homogeneous sample and calibration transfer method was proposed to solve the problem of detection inhomogeneous samples by NIRS.This work was focused on the tobacco leaf samples aspect.Three common calibration transfer methods, including Shenk′s patented algorithm (Shenk′s), piecewise direct standardization (PDS) and calibration transfer based on canonical correlation analysis (CTCCA), were used to construct two hybrid models of tobacco powder mixed with cut tobacco and tobacco powder mixed with tobacco flake samples to predict nicotine content in the samples of cut tobacco and tobacco flake.Experimental results showed that the hybrid model of adding some cut tobacco and tobacco flake samples to the powder model would get preferred prediction ability.Root mean square errors of cut tobacco and tobacco flake samples were reduced by 1.39% and 2.73%, respectively.This showed that the hybrid model was help for the improvement of the predicted results and the robustness of model.Moreover, CTCCA got the optimal performance between these three calibration transfer methods.Therefore, the scheme of building a hybrid model by NIRS homogeneous model and calibration transfer method to determinate the heterogeneous samples is feasible, which can accelerate the development of on-line near infrared spectroscopy technology and will provide reference for the share of near infrared spectral model.

Objective To investigate the effect of transcutaneous electric acupoint stimulation (TEAS)of Hegu-Neiguan-Houxi-Zhigou for preventing postoperative nausea and vomiting (PONV) in patients undergoing thoracoscopic lobectomy.Methods Sixty-four ASA physical status Ⅰ-Ⅲ pa-tients,aged 18-75 years,scheduled for elective thoracoscopic lobectomy were enrolled.Patients were randomly allocated into two groups (n =32):TEAS group (group E)and sham TEAS group (group C).TEAS at Hegu,Neiguan,Houxi and Zhigou was applied to patients before,during and after sur-gery in group E.The frequency was 2/100 Hz and the intensity was 2 times the threshold of sensation before and after surgery,and 30 mA during surgery.While sham TEAS with the intensity of sensory threshold was given to patients before and after surgery in group C.Patients in both groups received sufentanil,propofol and rocuronium for induction of anesthesia.Target controlled infusion of propofol and remifentanil was administered for maintenance of anesthesia.Patient-controlled intravenous anal-gesia (PCIA)was applied in both groups.No antiemetic was given to patients before and during sur-gery.Frequency of nausea or vomiting and any use of antiemetic for remediation were recorded after surgery.The postoperative Visual Analogue Scale (VAS)score and analgesic dosage were also docu-mented.Results Compared to group C,the incidence of nausea immediately after surgery,6 h after surgery,24 h after surgery and 48 h after surgery were significantly lower in group E.The incidence of vomiting 24 h after surgery was significantly lower in group E.The VAS score 6h and 24 h after surgery were significantly lower in group E.The analgesic dosage 48 h after surgery were reduced in group E.There were no significant differences regarding the use of antiemetic for remediation between the two groups.Conclusion TEAS at Hegu-Neiguan-Houxi-Zhigou is effective for preventing PONV in patients undergoing thoracoscopic lobectomy.

Objective To evaluate the clinical effect and safety of oxycodone hydrochloride in the anesthesia for percutaneous radiofrequency ablation (PRFA) in hepatocellular carcinoma.Methods Between March and December 2015,60 cases of hepatocellular carcinoma patients undergoing percutaneous radiofrequency ablation surgery in Peking University Cancer Hospital were randomly divided into three groups:oxycodone group (group Q),fentanyl group (group F) and dezocine group (group D),20 cases in each group.Respectively intravenously injection oxycodone 0.1 mg/kg,fentanyl 0.001 mg/kg,dezocine 0.1 mg/kg before surgery.After the surgeon completed puncture administer propofol to maintain anesthesia.Recorded mean arterial pressure (MAP),heart rate (HR),respiratory rate (RR),oxygen saturation (SpO2) changes in each group at entrance,beginning of radiofrequency ablation (T1),radiofrequency ablation began after 10 minutes (T2),the end of the surgical and awake.Observe the analgesia effect,respiratory depression,nausea,vomit and other complications.Postoperative pain scores were recorded.Using ANOVA,repeated measure variance analysis,SNK test,x2 test and other tests to evaluate the anesthetic effect indexes.Results The observation completed in all patients.Patients of three groups had no significant differences in general information.No significant difference between MAP,HR and SpO2 at each time points among the three groups.At the T1 time point (group Q:(11.7 ± 1.6)/min,group D:(12.1 ±l.7)/min,group F:(10.3 ±2.3)/min,F=5.068,P =0.009) and T2 time point (group Q:(11.9±1.3)/min,group D:(12.2±1.4)/min,group F:(1O.7±.1.3)/min,F=7.024,P=0.O02),RR in group F were lower than in group Q and group D.Pain visual analogue scores after waking (group Q:0.2 ±0.7,group D:O.3 ±0.7,group F:1.7 ± 1.5,F=12.981,P =0.000) and postoperative pain score of 1 hour (group Q:2.0 ± 0.9,group D:1.8 ± O.8,group F:4.3 ± 0.9,F =42.362,P =0.000) in the group Q and group D were significantly lower than in group F.The body movements in group Q and group D were significantly less than in group F (3 cases,3 cases,9 cases,x2 =6.400,P =0.041).Intraoperative respiratory depression in group Q and group D were lower than group F (3 cases,2 cases,9 cases,x2 =8.012,P =0.018).Conclusions Oxycodone hydrochloride can be used safely and effectively for radiofrequency ablation.It has favorable hemodynamic stability,lower incidence of respiratory depression,and advantage in terms of postoperative pain.

Objective To evaluate the clinical effect and safety of oxycodone hydrochloride in the anesthesia for percutaneous radiofrequency ablation (PRFA) in hepatocellular carcinoma.Methods Between March and December 2015,60 cases of hepatocellular carcinoma patients undergoing percutaneous radiofrequency ablation surgery in Peking University Cancer Hospital were randomly divided into three groups:oxycodone group (group Q),fentanyl group (group F) and dezocine group (group D),20 cases in each group.Respectively intravenously injection oxycodone 0.1 mg/kg,fentanyl 0.001 mg/kg,dezocine 0.1 mg/kg before surgery.After the surgeon completed puncture administer propofol to maintain anesthesia.Recorded mean arterial pressure (MAP),heart rate (HR),respiratory rate (RR),oxygen saturation (SpO2) changes in each group at entrance,beginning of radiofrequency ablation (T1),radiofrequency ablation began after 10 minutes (T2),the end of the surgical and awake.Observe the analgesia effect,respiratory depression,nausea,vomit and other complications.Postoperative pain scores were recorded.Using ANOVA,repeated measure variance analysis,SNK test,x2 test and other tests to evaluate the anesthetic effect indexes.Results The observation completed in all patients.Patients of three groups had no significant differences in general information.No significant difference between MAP,HR and SpO2 at each time points among the three groups.At the T1 time point (group Q:(11.7 ± 1.6)/min,group D:(12.1 ±l.7)/min,group F:(10.3 ±2.3)/min,F=5.068,P =0.009) and T2 time point (group Q:(11.9±1.3)/min,group D:(12.2±1.4)/min,group F:(1O.7±.1.3)/min,F=7.024,P=0.O02),RR in group F were lower than in group Q and group D.Pain visual analogue scores after waking (group Q:0.2 ±0.7,group D:O.3 ±0.7,group F:1.7 ± 1.5,F=12.981,P =0.000) and postoperative pain score of 1 hour (group Q:2.0 ± 0.9,group D:1.8 ± O.8,group F:4.3 ± 0.9,F =42.362,P =0.000) in the group Q and group D were significantly lower than in group F.The body movements in group Q and group D were significantly less than in group F (3 cases,3 cases,9 cases,x2 =6.400,P =0.041).Intraoperative respiratory depression in group Q and group D were lower than group F (3 cases,2 cases,9 cases,x2 =8.012,P =0.018).Conclusions Oxycodone hydrochloride can be used safely and effectively for radiofrequency ablation.It has favorable hemodynamic stability,lower incidence of respiratory depression,and advantage in terms of postoperative pain.

Coronary artery disease (CAD) is a complex human disease, involving multiple genes and their nonlinear interactions, which often act in a modular fashion. Genome-wide single nucleotide polymorphism (SNP) profiling provides an effective technique to unravel these underlying genetic interplays or their functional involvements for CAD. This study aimed to identify the susceptible pathways and modules for CAD based on SNP omics. First, the Wellcome Trust Case Control Consortium (WTCCC) SNP datasets of CAD and control samples were used to assess the joint effect of multiple genetic variants at the pathway level, using logistic kernel machine regression model. Then, an expanded genetic network was constructed by integrating statistical gene–gene interactions involved in these susceptible pathways with their protein–protein interaction (PPI) knowledge. Finally, risk functional modules were identified by decomposition of the network. Of 276 KEGG pathways analyzed, 6 pathways were found to have a significant effect on CAD. Other than glycerolipid metabolism, glycosaminoglycan biosynthesis, and cardiac muscle contraction pathways, three pathways related to other diseases were also revealed, including Alzheimer’s disease, non-alcoholic fatty liver disease, and Huntington’s disease. A genetic epistatic network of 95 genes was further constructed using the abovementioned integrative approach. Of 10 functional modules derived from the network, 6 have been annotated to phospholipase C activity and cell adhesion molecule binding, which also have known functional involvement in Alzheimer’s disease. These findings indicate an overlap of the underlying molecular mechanisms between CAD and Alzheimer’s disease, thus providing new insights into the molecular basis for CAD and its molecular relationships with other diseases.

Objective To compare the risk of tumor invasion and metastasis under paravertebral block (PVB) combined with general anesthesia versus general anesthesia in the patients undergoing radical resection for lung cancer performed via video-assisted thoracoscope in terms of plasma concentrations of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9).Methods Forty ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 30-64 yr,with body mass index of 18-25 kg/m2,scheduled for elective radical resection for lung cancer performed via video-assisted thoracoscope,were randomly divided into 2 groups (n =20 each) using a random number table:general anesthesia group (group G) and PVB combined with general anesthesia (group PG).PVB of T4-7 was performed successfully with local injection of 0.375％ ropivacaine 5 ml before induction of anesthesia.Double-lumen endotracheal tube was placed after induction of anesthesia,and the patients were mechanically ventilated.Anesthesia was maintained with inhalation of sevoflurane (end-tidal concentration 1％-2％),and intravenous infusion of remifentanil 0.2-0.3 μg · kg-1 · min-1,and intermittent intravenous boluses of atracurium.Before anesthesia and at 24 h after surgery,the venous blood samples were collected for measurement of plasma concentrations of VEGF and MMP-9.Results The plasma VEGF and MMP-9 concentrations were significantly lower after surgery in group PG than in group G.Conclusion PVB combined with general anesthesia significantly decreases the risk of tumor invasion and metastasis in the patients undergoing radical lung cancer resection performed via video-assisted thoracoscope in comparison to general anesthesia.