BACKGROUND: Oligodendrocyte precursor cell transplantation is one of the keys to the treatment of white matter damage in premature infants. At present, there is a lack of research on the comparison of oligodendrocyte precursor cells induced by neural stem cells derived from human fetal brain cultured in vitro in different vessels worldwide. OBJECTIVE: To observe the morphology of human oligodendrocyte progenitor cells and pre-oligodendrocytes in different cell culture vessels (6-well plates, 24-well plates and T25 flasks). METHODS: The 6-well plates, 24-well plates and T25 flasks were used as culture vessels to culture human oligodendrocyte progenitor cells and pre-oligodendrocytes. The characteristics of human oligodendrocyte progenitor cells were identified by immunofluorescence staining and flow cytometry. The morphology of cells was observed by an ordinary light microscope. Cell counts were performed according to cell morphology and statistical analysis was performed. RESULTS AND CONCLUSION: (1) The oligodendrocyte progenitor cell body was round and the bipolar protrusions were bead-like; the pre-oligodendrocyte protrusions were more than two poles, and did not bifurcate. (2) The ratios of oligodendrocyte progenitor cell morphology in the oligodontia lines were significantly higher in the 6-well plates than those in the 24-well plates and T25 flasks (P < 0.05), followed by T25 flasks and 24-well plates. Morphological ratios of pre-oligodendrocytes were significantly higher in the 24-well plates compared to the 6-well plates and T25 flasks (P < 0.01), followed by T25 flasks and 6-well plates. (3) The cells cultured in the 6-well plate had fewer dregs, and the morphology, vigor and growth were better than those of the other culture vessels. (4) According to morphological view, 6-well plates are more suitable for oligodendrocyte progenitor cell growth, and 24-well plates are more suitable for pre-oligodendrocytes growth.

Reflex tests are ordered when a particular test result indicates that additional testing should be performed according to the guidelines or the feedback process formulated by clinical consultation. The application scope of the reflex tests involves various subspecialties of laboratory medicine. The clinical application needs the support of qualified laboratory doctors, comprehensive information and financial system, clinical guidelines, and so on. Active application of reflex tests can promote the standardization of evidence-based medicine in clinical practice, save medical resources, and shorten the diagnosis and treatment time of patients.

Objective To observe the effect of Yijinjing Supporting Pile on the superficial microcirculation blood perfusion units (MBPU) on Du meridian in subjects with yang-deficiency constitution.Methods After the classification and determination of constitution according to the traditional Chinese medicine,19 volunteers (8 males and 11 females) with Yang-deficiency constitution were selected and provided with standard Yijinjing Supporting Pile training for 3 months.The laser Doppler flowmeter system was used to detect the superficial MBPU on the acupoints of Du meridian including Dazhui,Zhiyang and Mingmen,Results Before the training,the average MBPU value of Zhiyang was higher than Mingmen,but lower than Dazhui (P＜0.05,P＜0.01).After the training,the average MBPU value of all the acupoints increased significantly (P＜0.01).Conclusion Yijinjing Supporting Pilecan improve the superficial MBPU of Du meridian in subjects with Yang-deficiency constitution,and regulate the energy metabolism of the Du meridian.

Objective To explore nursing of the patients with acute myelogenous leukemia after upper abdominal cluster transplantation.Methods From March 2009 to August 2012,eight cases of upper abdominal organ cluster transplantation were performed in our center,among whom one suffered from acute myelogenous leukemia after operation.We summarized the clinical data and nursing methods of this case.Results The leukocytes,erythrocytes and platetets decreased 22 days after transplantation and the decreasing continued for more than four months.Four bone marrow biopsies were performed.During the first two biopsies,no definite cause of the blood cells decreasing was found,while the outcome of the third time showed that it was type M3 acute myeloid leukemia and genetic testing at the fourth time showed AML1/E-TO was positive.After diagnosis,the patient was shifted to the Department of Hematology and underwent further treatment.Conclusions Close observation,timely diagnosis and treatment,prevention of infection,bleeding and psychological nursing are the keys to promote patients recovery.

Objective To investigate the effect of intrathecal morphine preconditioning (ITMP) on the excitability of substantia gelatinosa (SG) neurons in the dorsal horn of the spinal cord in a rat model of myocardial ischemia-reperfusion (I/R).Methods Thirty-six adult male Sprague-Dawley rats,weighing 200-300 g,in which intrathecal catheters were successfully placed without complications,were randomly divided into 3 groups (n =12 each) using a random number table:sham operation group (group S),group I/R,and group ITMP.Myocardial I/R injury was produced by occlusion of the left anterior descending branch of the coronary artery for 30 min followed by 120 min reperfusion.In group ITMP,the rats received intrathecal morphine 3 μg/kg (10 μl) by three cycles of 5 min infusions interspersed with 5 min infusion-free periods starting from 30 min before ischemia,and the equal volume of normal saline was given instead of morphine in group I/R.At 10 min of reperfusion,6 rats randomly selected in each group were sacrificed,and the T2-6 segments of the spinal cords were acutely isolated to prepare spinal cord slices.The resting potential,threshold of action potential (APT),peak of action potential (APP) and action potential duration in SG neurons in the dorsal horn of spinal cord slices were determined using the whole-cell patch-clamp technique,and the number of action potentials evoked by currents of 40,60,80 and 100 pA was recorded.At 120 min of reperfusion,6 rats randomly selected in each group were sacrificed,and myocardial specimens were obtained for determination of myocardial infarct size (IS) and area at risk (AAR),and IS/AAR ratio was calculated.The expression of c-fos in the T2-5 dorsal horns of the spinal cords was detected by Western blot.Results Compared with group S,the IS/AAR ratio was significantly increased,the expression of c-fos was up-regulated,the number of action potentials in SG neurons in dorsal horns of spinal cord was increased,APT was decreased,and APP was increased in group I/R (P＜0.05).Compared with group I/R,the IS/AAR ratio was significantly decreased,the expression of c-fos was down-regulated,the number of action potentials in SG neurons in dorsal horns of spinal cord was decreased,APT was increased,and APP was decreased in group ITMP (P＜0.05).Conclusion The mechanism by which ITMP attenuates myocardial I/R injury is related to decrease in the excitability of SG neurons in the dorsal horn of the spinal cord and reduction of responses to nociceptive stimuli in rats.

Background and purpose:Ca2+plays a very important role in the maintenance of cell biological functions. The storage, release and uptake capacity of Ca2+ is controlled by endoplasmic reticulum (ER). Ca2+ homeo-stasis is essential for cellular energy metabolism and proper protein folding. This study aimed to investigate the effect of cytoplasmic Ca2+ on cisplatin induced ER stress-mediated autophagy in ovarian carcinoma SKOV3 and its underlying mechanism.Methods:The ovarian cancer SKOV3 was used as a study object. The experiment consisted of three parts:① To explore the possible relationship between cisplatin-induced ER stress and autophagy, SKOV3 cells were treated with cisplatin for 0, 6, 12 and 24 h, respectively;② To explore the possible relationship between ER stress induced Ca2+ effux and autophagy, SKOV3 cells were treated with cisplatin for 0, 9 and 12 h, respectively, and TG was used as a positive control;③ To explore the effects of blocking calcium effux on autophagy, SKOV3 cells were divided into control group, cisplatin group, TG group, BAPTA-AM group, cisplatin combined with BAPTA-AM group and TG com-bined with BAPTA-AM group. Western blot was used to detect the protein levels of GRP78 and LC3. Fluo-4 calcium lfuorescent probe was used to examine cytoplasmic Ca2+ levels. Confocal microscopy was used to detect LC3 level by immunolfurescence staining.Results:Compared to control group (0.679±0.011), GRP78 was signiifcantly accumulated at 6, 12 and 24 h after cisplatin treatment and reached the maximum value at 6 h (1.393±0.004,P=0.000). Similarly, compared to control group (0.038±0.000), LC3 puncta were clearly seen after cisplatin treatment and reached the maxi-mum value at 12 h (0.072±0.002,P=0.000). Using confocal microscopy, we found that cisplatin and TG increased LC3 punctate accumulation and cytoplasmic Ca2+ levels in a time-dependent manner. Immunolfuorescent method showed that treatment with cisplatin combined with BAPTA-AM or TG combined with BAPTA-AM increased LC3 punctate accumulation induced by cisplatin or TG. The results of Western blot showed that cisplatin combined with BAPTA-AM (0.071±0.001) or TG combined with BAPTA-AM (0.065±0.001) signiifcantly increased LC3Ⅱ/LC3Ⅰ ratio induced by cisplatin (0.039±0.000,P=0.000) or TG (0.035±0.001,P=0.000).Conclusion:Cisplatin induces intracellular ER stress and autophagy in SKOV3 cells, accompanied by increased cytoplasmic Ca2+ levels. Chelating cytoplasmic Ca2+enhanc-es cisplatin-induced autophagy.

Aim To investigate the effects of lentivirus mediated nerve growth factor ( NGF) gene silencing on pheochromocytoma cells ( PC12 ) and the possible mechanisms .Methods The NGF shRNA expression vector was constructed .PC12 cells were randomly divi-ded into five groups (n=3 each) as follows: negative control group ( NC ) , control lentivirus group ( LV CON) , lentivirus NGF shRNA1 group ( LV shNGF1 ) , lentivirus NGF shRNA2 group(LV shNGF2), lentivir-us NGF shRNA3 group(LV shNGF3).The cells in NC group were cultured in DMEM/HG and polybrene me-dium, while others were cultured in DMEM/HG, poly-brene and corresponding lentivirus medium .After the treatment, the infection efficiency was determined by fluorescent microscope .Relative expression of NGF , extracellular signal-regulated kinase ( ERK1/2 ) and p-ERK1/2 were assessed by Western blot .The expres-sion of NGF mRNA was analyzed by quantitative re-verse transcription polymerase chain reaction ( qRT-PCR) .The differentiation degree was valued according to the length of neuritis and max diameter of cells .The cell viability was detected by CCK-8.Results The in-fection efficiency in PC12 cells reached over 90%. Compared with NC group , the relative expression of NGF mRNA and NGF protein was significantly down-regulated ( P<0.05 ) .There was no difference in the expression of ERK1/2 protein and cell viability .The expression of p-ERK1/2 protein was markedly down-regulated in LV shNGF3 group ( P<0.01 ) .The cells morphology was changed , and the length of neuritis and max diameter of cells were strained in LV shNGF 3 group than those in NC group ( P<0.01 ) .Conclusion Lentivirus-mediated NGF gene silencing inhibits the differentiation of PC12 cells through suppressing the activation of ERK1/2.

Six DNA extraction methods and four DNA purification methods were compared and analyzed in this study to get higher quality DNA from the rhizospheric soil of Fritillaria thunbergii Miq.Results showed that higher purity DNA were harvested by pretreating the soil with 20 mmol/L EDTA(pH 7.5),then isolating soil DNA with CTAB-SDS-frozen-thawing,and further purified by agarose method.The recovery rate of this soil DNA was about 44.00 ?g/g ? 2.65 ?g/g soil,and they were qualified for the microbial diversity analysis in the rhizospheric soil of F.thunbergii Miq based on the 16S rDNA sequence.

An Aspergillus sp. strain F3 was isolated and identified from the rhizosphere soil of mangrove plant, Rhizophora stylosa Griff in Dongzhai harbor mangrove forest conservation in China. In this study, the effects of media salinity and pH on the mycelial biomass and the ability of producing antibacterial metabo- lites from this isolate were carefully analyzed. Results showed that this isolate can grow well on the SDA medium with higher salinity (3%~9%) and higher pH (8~10). Under the modified culturing conditions, this isolate can secret the antibacterial and antitumor metabolites. The extracts of acetic ether were about 448 mg/L of the fermentation broth. The antibacterial activities of the acetic ether extract were analyzed with bacteria and fungus. Results showed this extract can suppress the growth of Staphylococcus aureus、S. epi-dermidis、Sarcina lutea、Bacillus subtilis and Escherichia coli with MIC of 31.3 ?g/mL, 31.3 ?g/mL, 7.8 ?g/mL, 7.8 ?g/mL and 125.0 ?g/mL, respectively. It can also suppress the growth of Candida albicans with MIC of 125.0 ?g/mL. Further studies uncovered the cytotoxicity of this extract against the tumor cells, such as ECV304, Lovo and HepG2 with IC50 of 3.45 ?g/mL, 4.88 ?g/mL and 14.31 ?g/mL respectively.

Objective To investigate the nursing points of pancreatic leakage after upper abdominal multiple organ transplantation. Methods A retrospective study was conducted on the nursing experience of two patients with end-stage liver disease and type 2 diabetes mellitus who were complicated with pancreatic leakage after upper abdominal multiple organ transplantation from March 2009 to July 2015. Results The blood glucose of these patients returned to normal level within 1 week after operation and insulin was discontinued. Pancreatic leakage was occurred in the two patients at 14 and 21 days after operation, respectively. They were both successfully discharged after active treatments and nursing cares including completely drainage, the application of drugs that inhibited the secretion of pancreatic enzymes and digestive tract glands, strengthening infection control, nutritional support and other conservative treatments. Conclusions It is the key to improve the recovery of pancreatic leakage after upper abdominal multiple organ transplantation with careful observation of abdominal signs and abdominal drainage tube, accurate use of somatostatin, nutritional support, maintenance of water and electrolyte balance, and psychological intervention.