Autophagy is a vacuolar process of cytoplasmic degradation by lysosome which ubiquitously occurring in all eukaryotic cells. The researches of autophagy have made great progress with the development of the yeast model and genetic technology. This review will summarize the determination of autophagy, its relationship with apoptosis and its role in the tumor treatment in order to give a comprehensive understanding of the function of autophagy.

Objective Near-infrared (NIR) reflectance spectroscopy was used to develop a fast quality assessment method by simultaneous material identification and moisture quantification of Radix Ginseng Rubra (RGR). Methods The sample was identified by the comparison of its spectrum with a standard NIR spectral library. Similarity measurement was used as the discriminating parameter. The moisture content of sample was quantified by a partial least square (PLS) calibration model, correlative spectrum calculation was used for wavelength selection and multiplicative signal correction (MSC) was applied for pretreatment in the calibration model. Primary reference data were obtained using the traditional loss on drying (LOD) method. Results The NIR library can distinguish RGR from the counterfeit successfully. The optimized eight-factor PLS calibration model of NIR spectra has a high correlation coefficient (R=(0.999 7).) Conclusion The proposed method is rapid, accurate and can be used routinely in the traditional Chinese materia medica manufacturers for quality control of raw materials.

Objective To compare the ventilatory effects between three-way laryngeal mask airway (TLMA)and tracheal catheter (TC) on hemodynamics, respiratory function and stress responses on patients during bronchoalveolar lavage (BAL). Method Forty patients scheduled for BAL under general anesthesia were divided (stratified sampling) into either TLMA group (group T,n = 20) or TC group (group C, n = 20) according to the stratified sampling principle. SpO2, SBP, DBP and HR were measured in 5 min after entering the operating theater (To), just before inserting TLMA or TC(T1), immediately after inserting TLMA or TC(T2) ,3 min(T3), 5 min(T4), 10 min(T5)after mechanical ventilation, 10 min(T6),20 min(T7), 30 min(T8)during the course of BAL,immediately after extubating TLMA or TC (T9)and 3 min after extubating TLMA or TC (T10). The tidal volume (VT), peak inspiratory airway pressure (Ppeak) and end expiratory CO2 pressure(PETCO2)were recorded at T2,T4,T6,T7, T8, T10. The venous blood samples were taken at T0, T2, T3, T4, T6, T9, T10 for the measurements of epinephrine(AE), norepinephrine(NE)and dopamine (DA) levels with high performance liquid chromatography.Data were dealt with SPSS version 10.0 statistic software. The variables of hemodynamics and stress responses were analyzed with ANOVA of repeating test data. P ＜ 0.05 means the difference in statistical significance. Results In group C, SBP, DBP and HR were significantly higher than those in group T at T2 ,T3 ,T9 (P ＜ 0.05). In group C, the levels of Ppeak were significantly higher than those in group T at T6 ,T7 ,T8 (P ＜ 0.05), and the concentrations of AE, NE and DA were also significantly higher in group C than those in group T at T2, T3 and T9 (P ＜0.05). Conclusions Ventilation with TLMA in patients during BAL is better than TC in respects of keeping stable ventilation, stable hemodynamics and producing less stress responses.

Objective To investigate the value of 18F-FDG,11C-MET and 11C-CHO PET/CT in the differentiation of C6 glioma from different kinds of inflammation in experimental rat models.Methods (1) A total of 48 male SD rats were randomly divided into 6 groups by the random number table:group 1 and 2 consisted of 8 rats bearing both C6 glioma and turpentine oil-induced acute inflammation;group 3 and 4 consisted of 8 rats bearing both C6 glioma and turpentine oil-induced chronic inflammation;group 5 and 6 consisted of 8 rats bearing both C6 glioma and BCG-induced granuloma.(2) 18F-FDG and 11C-MET PET/CT were performed on rats of group 1,3 and 5;18F-FDG and 11C-CHO PET/CT were performed on rats of group 2,4 and 6.The lesion-to-muscle ratios and tumor selectivity index (SI) were calculated.(3)After the PET/CT imaging,the lesions were excised.Immunohistochemical staining was used to demonstrate the situation of Glut-1,HIF-1α and CD98.(4)Two-sample t test,Nemenyi test and nonparametric Kruskal-Wallis H test were used for statistical analyses.Results (1) 18 F-FDG and 11 C-MET uptake in C6 glioma were higher than those in different inflammatory tissues(t--1.425-3.901,all P＜0.05).The 11 C-CHO uptake among different lesions were not significant (t =0.031-3.901,all P＞0.05).In group 1 and 5 models,SIMET(4.22±2.96 and 4.89±2.08) was significantly higher than SIFDG(1.77±0.86 and 1.72±0.77;t =2.717and 2.490,both P＜0.05);but iu group 3 models,SIMET(3.84±2.71) was not significantly higher than SIFDG(2.28± 1.14;t =2.082,P＞0.05).(2) Immunohistochemical study showed that there were significant differences in the expression of HIF-1 α,CD98 among different lesions (H =17.810,26.540,both P ＜ 0.05),and no significances of expression of Glut-1 among different lesions (H=5.940,P＞0.05).Nemenyi test showed that there was significant difference for CD98 expression between C6 glioma and acute inflammation,C6 glioma and granuloma (x2=5.504,9.345,both P＜O.05),and for HIF-1α and CD98 expression between C6 glioma and chronic inflammation (x2 =-5.938,2.128,both P＜0.05).Conclusions Compared with 18F-FDG and 11 C-CHO,11 C-MET has better tumor specificity.11 C-CHO PET/CT is not suitable for the differentiation of tumor and inflammation because of its lowest specificity.

OBJECTIVE: To investigate the mechanism of Chinese herbal recipe Weichang'an (WCA) in inducing cell apoptosis of human gastric cancer grafted onto nude mice. METHODS: The high performance liquid chromatography was used for monitoring the stability of WCA. A human gastric cancer cell line SGC-7901 grafted in nude mouse was used as the animal model. The mice were divided into untreated group and two experimental groups. Animals in the two experimental groups received either WCA over a 34-day period or 5-fluorouracil (5-FU) over a 6-day period starting at the 8th day after grafting. Animals in the untreated group received normal saline on an identical schedule. Animals were killed 41 days after being grafted. To assess the effect of the treatment on tumor, the tumor weight was determined by the electron balance immediately after the animals were killed. SP immunohistochemical method was used to detect the expression of proliferating cell nuclear antigen (PCNA) in grafts. Apoptotic indices (AI) of the tumor cells were examined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method. SP method was also used to detect the expressions of cleaved caspase-3, caspase-8 and caspase-9. SYBR green dye I real-time quantitative polymerase chain reaction (real-time quantitative [corrected] PCR) was used to assess the related gene alterations in mRNA level. The expressions of phospho-Stat3 (Tyr705) and bcl-2 proteins were detected by using SP method. RESULTS: Compared with the untreated group, tumor growth was significantly inhibited by treatment of WCA or 5-FU (P<0.01, respectively). The tumor inhibition rate in the WCA-treated group was 48.70% and that in the 5-FU-treated group was 60.10%. The average labeling index (LI) for PCNA in the WCA-treated group and 5-FU-treated group was significantly decreased as compared with that in the untreated group, respectively. The AI of human gastric cancer grafted in the nude mice detected by using TUNEL method was significantly increased to (9.72+/-4.51)% in the WCA-treated group, while it was (2.45+/-1.37)% in the untreated group. 5-FU-treated group was also found a significantly increased AI compared with the untreated group. The expressions of cleaved caspase-3 and caspase-9 in the WCA-treated group and 5-FU-treated group were significantly increased as compared with those in the untreated group. But caspase-8 showed no significant alteration either in the WCA-treated group or in the 5-FU-treated group. The expression levels of Stat3 (2(-)delta delta C(T))=0.16) and bcl-2 (2(-)delta delta C(T))=0.10) detected by using real-time quantitative [corrected] PCR were lower in the WCA-treated group than those in the untreated group. The expressions of phospho-Stat3 (Tyr705) and bcl-2 in the WCA-treated group were significantly decreased as compared with those in the untreated group. CONCLUSIONS: Chinese herbal recipe WCA can inhibit gastric cancer cell SGC-7901 growth in vivo, induce gastric cancer cell apoptosis and suppress the cell proliferation. WCA induces apoptosis through the caspase-9 and caspase-3 pathway in vivo. Its mechanism might be involved in the down-regulation of Stat3 and bcl-2 genes.

OBJECTIVE: To evaluate the growth-inhibiting and anti-metastasis effects of Weichang'an Decoction (WCAD) on orthotopic transplant nude mouse model of human gastric cancer. METHODS: Forty-one nude mice were implanted with SGC-7901 cells at orthotopic site, whereas 25 were implanted with SGC-7901 cells subcutaneously. Then the nude mice in each transplantation model were divided into the same three groups which were WCAD-treated group with WCAD 0.5 ml/d taken orally, 5-FU-treated group with 5-FU 50 mg/kg intraperitoneally injected weekly and untreated control group with physiological saline 0.5 ml/d taken orally. The growth-inhibiting rates of transplant tumors were detected and the metastatic lesions were examined in the orthotopic transplant mouse model while PCNA-positive rate and apoptotic index (AI) were observed in the subcutaneous transplant mouse model. RESULTS: The growth-inhibiting rates in the WCAD-treated and 5-FU-treated groups of orthotopic transplant mouse model were 40.82% and 37.92% respectively whereas those of subcutaneous transplant mouse model were 48.70% and 60.10%. The incidence rates of metastasis in perigastric lymph notes, lymph nodes in the porta hepatis, liver, diaphragm and peritoneum in the WCAD-treated and 5-FU-treated groups were lower than those in the untreated control group, and the total metastasis rates in the WCAD-treated, 5-FU-treated and the untreated control groups were 30.77%, 28.57% and 71.43% respectively with significant differences (P<0.05). The total PCNA-positive rates in the WCAD-treated and 5-FU-treated groups were obviously lower than that in the untreated control group (P<0.05 or P<0.01) while the AI was higher than that in the untreated control group (P<0.01). The growth-inhibiting rate, total PCNA-positive rate and total metastasis rate in the WCAD-treated group had no significant differences as compared with those in the 5-FU-treated group, but the AI in the WCAD-treated group was significantly higher than that in the 5-FU-treated group (P<0.05). CONCLUSION: WCAD has the inhibiting effects on tumor growth and metastasis of gastric cancer which is orthotopic implanted onto nude mice. This effect may be obtained by proliferation suppression and apoptosis induction in cancer.

Three-way laryngeal mask airway (tLMA) was used in 31 patients aged 4-68 yr, weighing 10- 79 kg undergoing tracheal foreign body removal under general anesthesia. Anesthesia was induced with propofol 3 mg/kg, vecuronium 0.12 mg/kg and remifentanil 0.4 μg/kg. tLMA was inserted. The patients were mechanically ventilated. Anesthesia was maintained with iv infusion of propofol 2 mg . Kg-1 ? H-1, vecuronium 0.08 mg·kg-1·h-1 and remifentanil 0.15 μg·kg-1 ·min-1 . Radial artery was cannulated for BP monitoring and blood sampling. The operation time was 6-34 min and mechanical ventilation time 19-45 min. There was no significant change in SP, DP, HR, VT, Ppeak and Ppeak CO, during operation as compared with the baseline values before anesthesia. SpO2 was significantly increased at T2-6. PCO2, PO2 and O2sat were obviously improved after tLMA was used. All the patients emerged bom anesthesia within 30 min after operation. No aspiration, obvious gastrointestinal inflation, and pharyngeal and laryngeal edema and injury occurred. Mild agitation occurred in a short time during the recovery period in one patient. No complication occurred.

Objective:To observe the effect of Hippocampus kelloggi on GRP-78/PERK/ATF-4 signal pathway and explore its mechanism on improving spinal cord injury. Methods:A total of 36 SD rats were randomly divided into sham operation group, model group and hippocampus group with 12 rats in each group. Only laminectomy was performed in the sham operation group. The spinal cord injury model was prepared in the model group and hippocampus group. Rats in the hippocampus group were given 10 ml/kg Hippocampus kelloggi extract by gavage for 14 days. Basso Beattie Bresnahan (BBB) score was used to evaluate the motor function of the limbs. The neuron morphology was observed by Nissl staining. The expression of GRP-78, p-PERK and ATF-4 proteins were detected by Western blot, the expression of GRP-78 and ATF-4 mRNAs was detected by qPCR, Caspase-3 and Caspase-12 were detected by ELISA, and the apoptosis was detected by TUNEL. Results:Compared with the model group, the BBB score of hippocampal group increased on the 7th, 9th, 11th and 14th day after operation ( P<0.05). For hippocampus group, the relative expression of GRP-78 (0.49 ± 0.06 vs. 0.74 ± 0.03), p-PERK (0.63 ± 0.04 vs. 0.81 ± 0.06) and ATF-4 (0.51 ± 0.06 vs. 0.69 ± 0.05) protein were significantly decreased ( P<0.05), GRP-78 mRNA (0.54 ± 0.05 vs. 0.63 ± 0.06) and ATF-4 mRNA (0.61 ± 0.06 vs. 0.78 ± 0.04) were significantly decreased ( P<0.05), the content of Caspase-3 and caspase-12 were significantly decreased ( P<0.05), and the apoptosis rate of hippocampal group was significantly decreased ( P<0.05). Conclusion:Hippocampus kelloggi can regulate the stress response of the endoplasmic reticulum after spinal cord injury by inhibiting GRP-78/PERK/ATF-4 signaling pathway to promote the repair of neurons.

Objective:To investigate the clinical efficacy of ex vivo liver resection and autotransplantation (ELRA) for liver complex space-occupying lesions.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 50 patients with liver complex space-occupying lesions who underwent ELRA in the First Hospital Affiliated to Army Medical University between June 2009 and May 2017 were collected. There were 36 males and 14 females, aged from 13 to 69 years, with a median age of 51 years. All patients underwent ELRA. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted by outpatient examination or telephone interview. Patients were followed up according to the individual follow-up plan in the first 6 months after discharge, and then once every 3 to 6 months to detect tumor recurrence and survival up to May 2019. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represented as M (range). Count data were expressed as absolute numbers or percentages. The Kaplan-Meier method was used to calculate survival rates and draw survival curves, and Log-rank test was used for survival analysis. Results:(1) Surgical situations: all the 50 patients underwent ELRA successfully, and postoperative pathological examination showed the R 0 resection rate was 100%(50/50). The operation time of the 50 patients were (630±186)minutes, of which 9 patients with liver benign occupation had the operation time of (684±168)minutes and 41 patients with liver malignant tumor had the operation time of (618±190)minutes. The operation time of temporary reconstruction of inferior vena cava and portacaval shunt, time of anhepatic phase, volume of intraoperative blood loss of the 50 patients were (35±9)minutes, (256±71)minutes, 2 000 mL(range, 400-10 000 mL), respectively. The remnant liver mass to standard liver mass ratio of the 50 patients was 65%±16%, of which 9 patients with liver benign occupation had the remnant liver mass to standard liver mass ratio of 63%±14% and 41 patients with liver malignant tumor had the remnant liver mass to standard liver mass ratio of 65%±17%. Of the 50 patients, 35 had vascular invasion (7 cases with liver benign occupation, 28 cases with liver malignant tumor), of which 24 (6 cases with liver benign occupation, 18 cases with liver malignant tumor) underwent in vitro vascular reconstruction, 12 (5 cases with liver benign occupation, 7 cases with liver malignant tumor) had bile duct invasion and underwent choledochojejunostomy due to the inability of the common bile duct to anastomose the ends. Two cases with liver metastasis of gastric cancer, one case with liver metastasis of colon cancer and one case with liver metastasis of pancreatic cancer underwent radical gastrectomy, radical resection of colon cancer, and pancreaticoduodenectomy, respectively. (2) Postoperative situations: the duration of postoperative hospital stay of the 50 patients were 25 days (range, 11-169 days). Of the 50 patients, 12 had pleural effusion who were treated with pleural puncture drainage, 10 had bile leakage who were treated with abdominal puncture drainage, 3 had bile duct anastomotic leakage who were treated with endoscopic nasobiliary drainage or biliary stent implantation, 6 underwent reoperation among which 4 underwent exploratory laparotomy due to abdominal hemorrhage, 1 underwent portal vein reconstruction due to abdominal hemorrhage combined with portal vein thrombosis, and 1 underwent salvage liver transplantation due to liver failure. Nine of the 50 patients died within 90 days after surgery, all of whom had liver malignant tumor. Among them, 3 died of multi-organ dysfunction syndrome caused by severe infection, 3 died of acute liver failure, 2 died of abdominal hemorrhage and 1 died pulmonary embolism. (3) Follow-up: all the 50 patients were followed up for 1 to 119 months. The overall survival time, 1-, 3-, 5-year overall and tumor-free survival rates of the 50 patients after operation were 17 months (range, 1-119 months), 68.0%, 45.9%, 41.1% and 41.9%, 33.4%, 30.8%, respectively. The overall survival time, 1-, 3-, 5-year overall and tumor-free survival rates of the 9 patients who with liver benign occupation after operation were 68 months (range, 10-114 months), 88.9%, 88.9%, 88.9% and 88.9%, 88.9%, 88.9%, respectively. The overall survival time, 1-, 3-, 5-year overall and tumor-free survival rates of the 41 patients who with liver malignant tumor after operation were 15 months (range, 1-119 months), 63.4%, 36.6%, 31.0% and 31.5%, 21.0%, 18.0%, respectively. There were significant differences in the overall and tumor-free survival rates between patients who with liver benign occupation and patients who with liver malignant tumor ( χ2=7.626, 11.766, P<0.05). Conclusions:ELRA can be applied in the treatment of liver complex space-occupying lesions. The selection criteria of patients with liver malignant tumor should be more rigorous to reduce perioperative mortality.

OBJECTIVETo build a protocol of separation and induction of megakaryocytes derived from cord blood mononuclear cells.

METHODSRed blood cells were precipitated by hydroxyethyl starch (HES). Mononuclear cells were obtained by density gradient centrifugation with Ficoll. The inducing efficiencies of megakaryocytes by using of different cytokine cocktails and culture media were analyzed.

RESULTSThe best choice for erythrocyte sedimentation and high efficiency of nucleated cells retrieving were obtained by using of 1.5% HES. The isolated cord blood mononuclear cells were cultured with domestic serum-free medium supplemented with 116t (IL-11, IL-6, TPO), st36(SCF, TPO, IL-3, IL-6), pt36 (PDGF,TPO,IL-3,IL-6) or pst36 for 7 days. St36 group (50 ng/ml SCF, 50 ng/ml TPO, 20 ng/ml IL-3 and 50 ng/ml IL-6) yielded the most CD41/CD61 positive [(6.79±1.97)×10⁴]. The cell viability [(82.85 ± 0.64)%] of st36 group by using of imported serum-free medium was better than [(60.90±6.93)%] that in domestic medium on day 7 after induction, and CD41/CD61 positive cells count [(18.60±1.97)×10⁴] were more than domestic serum-free medium group. Therefore, we chose imported serum-free medium containing st36 to induce cord blood mononuclear cells. After a prolonged culture, the total cell numbers increased accompanied with an elevated percentage of CD41/CD61 positive cells, which reached (54.27 ± 6.31)% on day 14. Wright-Giemsa staining showed that different phase cells, such as megakaryoblast, promegakaryocyte and granular megakaryocyte, occurred after 10 days'culture. Clone forming unit-megakarocytes (CFU-MK) assay showed that the colonies count increased with the prolonged incubation. CFU-MK colonies were [1 236.0±32.9] on day 14, which was higher than that in medium without induction (P<0.01). Platelets from megakaryocytes showed agglutination function after 10 days'culture.

CONCLUSION1.5% HES was the best solution to precipitate erythrocytes. The combination of an imported serum-free medium with IL-3, IL-6, SCF and TPO showed better induction efficiency than domestic medium or other cytokine cocktails. Meanwhile, induced megakaryocytes produced functional platelets.

Cell Culture Techniques ; Cell Differentiation ; Cell Division ; Cell Separation ; methods ; Cells, Cultured ; Culture Media, Serum-Free ; Fetal Blood ; cytology ; Humans ; Megakaryocyte Progenitor Cells ; cytology