Objective Through studying the apoptosis induced by stichopus japonicus acid mucopoly saccharide in the hepatocellular carcinoma cell line HepG2 in vitro, analysing the expression of Bcl-2 and nm-23in HepG2, to provide the theory foundation and its feasibility on whether it can be used for the chemotherapy of hepetocellular carcinoma. Methods The cells of HepG2 were cultured in vitro and treated with SJAMP at different doses(0.25,0. 5,1.0,2.0,4.0 g/L). MTT was used to observe the inhibitory effects of SJAMP on cell growth, Western blotting was used to detect apoptosis, and the apoptosis related change of expression of protein Bcl-2 and nm23-H1. Results (1) MTT identified that SJAMP produced an obvious time-and-dose-dependent inhibitory effect on the HlepG2 cells. (2) Western blot showed that SJAMP could induce the apoptosis of HepG2 cells through changing the expression of the protein of Bcl-2 and nn23-H1 (P<0.05). Conclusion (1)SJAMP produced obvious inhibitory effects on HepG2 cells and induce HepG2 apoptosis. (2)SJAMP can enduce the anti-tumor function in the method of changing the expression of protein Bcl-2 and nm23-H1.

Objective To study the influence of pre-injection of donor apoptotic cells in the survival of islet grafts and the function of T lymphocytes in the peripheral blood. Methods The donor apoptotic cells and necrotic cells were ob-tained respectively by X-irradiation from electron linear accelerator and a heat-shock procedure (water bath box 56℃, 1 h). The diabetic rats for islet transplantation (n=42) were induced by a single intraperitoneal injection of streptozotocin (STZ), then were randomly divided into four groups:rats were injected by physiological saline group (n=9), normal cells group (n=12), apoptotic donor cell group (n=12) and necrotic donor cell group (n=9). On the seventh day, each group received islet transplantation under the renal capsule. The blood glucose level was detected to reflect the survival of the islets. The periph-eral blood samples of three rats in each group were obtained at different observation times. The proliferative activity of T lym-phocytes was determined by MTT method. The levels of cytokines interferon (IFN)-γ, interleukin (IL)-10 in peripheral blood were measured by Luminex 100 Integrated System, and transforming growth factor (TGF)-β1 by ELISA respectively at 0 d, 1 week, 2 weeks and after rejection. Results The survival time of islets was significantly prolonged by the pre-intervention of apoptotic cells, and the proliferative activity of T lymphocytes stimulated by ConA was inhibited. Meanwhile, the extent of the increased level of IFN-γwas inhibited significantly at 1 week and 2 weeks after transplantation (P<0.05), the levels of IL-10 and TGF-β1 were significantly increased before transplantation, 1 week and 2 weeks after transplantation (P<0.05). Conclusion Our results demonstrated that the pre-treatment of donor apoptotic cells can regulate the recipient’s immune reactive state by inhibiting the proliferative activity of T lymphocytes and changing the levels of cytokines from different sub-sets of T lymphocytes, and finally resulted in the prolonging of the survival of islet grafts.

ObjectiveTo evaluate gallbladder conserving gallstone removal and polyps resection using combination laparoscopy,hard gallbladder endoscopy and soft choledochoscopy.MethodsClinical data of 122 patients with cholecystolithiasis or polyps undergoing removal of calculus (polyps) and preservation of gallbladder were analyzed retrospectively.ResultsGallstones in 56 patients and polyps in 24 cases was removed or resected successfully by laparoscopy and hard gallbladder endoscopy; In the remaining 34 cases stones were completely removed by combination soft choledochoscopy; 8 cases were converted to laparoscopic cholecystectomy.Romoved stone was single in 25 cases and multiple in 65 cases,with the number ranging from 1to 52,the diameter of stone ranged from 0.2 cm to 3.2 cm.In the 24 gallbladder polyps,7 cases were single,17 cases were multiple,the diameter of polyp ranged from 0.8cm to 1.2 cm.The operation time was 40-125 (78) min. The mean hospitalization was 4 days. No intraoperative and postoperative complications occurred.All patients were followed up for 1year.Gallstones recurred in 3 cases,and the recurrence rate was 3.06％. ConclusionsLaparoscopy combined with hard gallbladder endoscopy and soft choledochoscopy for removing calculi (polyp) and conserving gallbladder is safe and feasible.

Objective To investigate the effects of environmental hypothermia on hemodynamics and oxygen metabolism in unanesthetized swine model of hemorrhagic shock.Methods A total of 16 Bama pigs provided by animal experiment centre of the General Hospital of Shenyang Military Command were randomized into two groups ( n =8,each):ambient temperature (A) and hypothermia ( H ).Venous blood (30 mL/kg) was continously withdrawn over 15 minutes to establish hemorrhagic shock model.Core temperature,heart rate,mean arterial pressure,central venous pressure,cardiac output,saturation of mixed venous blood and blood gas analysis were recorded at the baseline and different hemorrhagic shock time.DO2I and VO2I,and the O2 extraction ratio (O2ER) were calculated.Results Core body temperature in group A decreased slightly after hemorrhagic shock model had established and environmental hypothermia resulted in more reduction in core body temperature.The mortality rate was significantly higher in group H (50％) than in group A (0％) (P ＜0.05).DO2I and VO2I decreased significantly after hemorrhage.No difference was found in hemodynamics,DO2I and VO2I between group A and group H,but the difference of pH,lactic acid and O2ER were significant between the two groups (P ＜ 0.05 ).conclusions Environmental hypothermia aggravated the disorder of oxygen metabolism after hemorrhagic shock,which was associated with poor prognosis.

Objective To compare the anesthetic effects of interscalene brachial plexus combined with ulnar nerve and axillary brachial plexus block guided by nerve stimulator. Methods Eighty patients belonging to ASA ⅠorⅡ and undergoing replantation of severed palm or wrist were divided randomly into 2 groups, Each group had 40 patients. Nerve stimulator guided nerve block. Patients in groupⅠreceived interscalene brachial plexus combined with ulnar nerve block, and those in groupⅡreceived axillary brachial plexus block. The onset time, hold time, tourniquet tolerance of radial nerve, median nerve and ulnar nerve of two groups was recorded. The phrenic nerve block, Horner′s syndrome and recurrent laryngeal nerve block was compared between two groups. Results The onset time of radial nerve, median nerve and ulnar nerve in group Ⅰwas (5.13 ± 0.76), (7.13 ± 1.04), (3.23 ± 0.62) min , in group Ⅱ was (9.23 ± 1.61), (12.35 ± 1.76), (8.83 ± 1.13) min, and there were significant differences (P<0.05). The excellent rates of sensory block of radial nerve, median nerve and ulnar nerve in group Ⅰ were 90.0%(36/40), 85.0%(34/40), 97.5%(39/40), in group Ⅱ were 72.5%(29/40), 65.0%(26/40), 70.0%(28/40), and there were significant differences (P<0.05). The full rates of motor block of radial nerve, median nerve and ulnar nerve in groupⅠwere 75.0%(30/40), 37.5%(27/40), 80.0%(32/40), in groupⅡ were 47.5%(19/40), 40.0%(16/40), 45.0%(18/40), and there were significant differences (P < 0.05). The tourniquet tolerance rate in group Ⅰwas significantly higher than that in groupⅡ:90.0%(36/40) vs. 62.5%(25/40) , P<0.05. In groupⅠ, phrenic nerve block occurred in 2 patients, and Horner syndrome occurred in 1 patient. None had laryngeal recurrent nerve block in both group. Conclusions The interscalene brachial plexus combined with ulnar nerve block guided by nerve stimulator is more suitable for a long time microsurgery of the palm or wrist, because it takes action faster, has better sensory and motor block effects, improves the rate of tourniquet tolerance without increasing untoward reaction.

To establish human beta-defensin-3 gene transgenic cell lines as competent donor cells for the production of transgenic animals using somatic cell nuclear transfer (SCNT). Firstly, we obtained human beta-defensin-3 by RT-PCR from human placenta, and subsequently inserted the fragment hBD into the corresponding site of the plasmid pBCP. Then we moved the combined fragment BCD (including 5' and 3' regulating region of beta-casein and hBD) into the corresponding site of the plasmid pEGFP-C1. Finally we successfully constructed mammary-specific expression vector pEBCD. We transected pEBCD into Holstein Fetal fibroblast cells by Lipofectamine TM-2000 and selected in medium with G418 for three to four weeks. We identified G418 resistant transfectants by PCR, RT-PCR and EGFP detection. Our results indicated that human beta-defensin-3 gene stably was integrated into the open region of the chromatin in G418 resistant fibroblast cells. Meanwhile we identified the expression of human beta-defensin-3 in the supernatant of stable transfected mammary epithelial cells by Western blotting. This study may provide competent transgenic donor cells for the production of transgenic animals by SCNT and improve the efficiency of transgenic cloning.
Animals ; Animals, Genetically Modified ; Caseins ; genetics ; Cattle ; Epithelial Cells ; metabolism ; Genes, erbB-1 ; genetics ; Genetic Vectors ; genetics ; Humans ; Mammary Glands, Animal ; cytology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; beta-Defensins ; biosynthesis ; genetics

To discuss the effect of co-culture on the quality of mouse blastocysts and their epigenetic modification. We divided mouse zygotes into three co-culture experiment groups : with granular cells (group I ), oviduct epithelium cells (group II) and oviduct tissue (group III). Meanwhile, we set up control A (cultured in vitro, only KSOM (KCl+ simplex optimized medium)) and control B (cultured in vivo). Then we compared cleavage rate and blastocyst rate among different groups. After that we evaluated the quality of blastocysts by using ICM/TE (Inner cell mass/Trophectoderm cells) ratio via staining with propidium iodide and Hoechest333258, and analyzed the level of genome methylation and histone acetylation by immunofluorescence. Compared with the control group A, the co-culture groups had increased cleavage rate and blastocyst rate (P < 0.05), blastocyst cells and the ICM/TE ratio of co-culture groups were higher (P < 0.05), the level of genome methylation and histone acetylation had no significant difference between groups in vitro (P > 0.05), but the level of genome methylation in vivo was significantly higher than that of in vitro (P < 0.05). The co-culture methods can successfully promote the development rate of embryos in vitro, and improve the quality of the blastocyst. However, the methods have drawbacks in changing the abnormal genome methylation with in vitro culture.
Animals ; Blastocyst ; cytology ; Coculture Techniques ; DNA Methylation ; Epigenesis, Genetic ; genetics ; Epithelial Cells ; cytology ; Fallopian Tubes ; cytology ; Female ; Male ; Mice

Objective:To evaluate the curative effect of non-surgical treatment of acute upper gastrointestinal perforation and analyze the risk factors.Methods:We retrospectively reviewed medical records of patients who were diagnosed with acute upper gastrointestinal perforation from Jan 2016 to Dec 2018 in Liaocheng People's Hospital. At first, all patients were put on non-surgical treatment. According to whether or not converted to surgery, they were divided into non-surgical treatment group (163 cases) and surgery group (29 cases). Univariate analyses and multivariate analyses were conducted.Results:192 patients with acute upper gastrointestinal perforation were cured without serious complications and death. The non-surgical treatment efficiency was 84.9%. The onset time ( OR=0.238, P=0.046), heart rate ( OR=1.043, P=0.004), serum albumin ( OR=0.869, P=0.002) are independent risk factors. Conclusion:Non-surgical treatment of acute upper gastrointestinal perforation is safe and effective. Onset time, heart rate and serum albumin are independent risk factors. In patients when time of onse t>12h, heart rate >100 beats/min, hypoalbuminemia, and high level of procalcitonin , conversion to surgery should be considered.

Multi-modal brain-computer interface and multi-modal brain function imaging are developing trends for the present and future. Aiming at multi-modal brain-computer interface based on electroencephalogram-near infrared spectroscopy (EEG-NIRS) and in order to simultaneously acquire the brain activity of motor area, an acquisition helmet by NIRS combined with EEG was designed and verified by the experiment. According to the 10-20 system or 10-20 extended system, the diameter and spacing of NIRS probe and EEG electrode, NIRS probes were aligned with C3 and C4 as the reference electrodes, and NIRS probes were placed in the middle position between EEG electrodes to simultaneously measure variations of NIRS and the corresponding variation of EEG in the same functional brain area. The clamp holder and near infrared probe were coupled by tightening a screw. To verify the feasibility and effectiveness of the multi-modal EEG-NIRS helmet, NIRS and EEG signals were collected from six healthy subjects during six mental tasks involving the right hand clenching force and speed motor imagery. These signals may reflect brain activity related to hand clenching force and speed motor imagery in a certain extent. The experiment showed that the EEG-NIRS helmet designed in the paper was feasible and effective. It not only could provide support for the multi-modal motor imagery brain-computer interface based on EEG-NIRS, but also was expected to provide support for multi-modal brain functional imaging based on EEG-NIRS.