Objective To investigate the effective mechanism, time and rational dose of radiotherapy of epileptic rat. Methods To make rat model of epilepsy, and give 0, 24,12 or 6Gy X ray irradiation, and then detect the amino-acid contents in frontal lobe of those rats in 1 hour, 24 hours or 1 week after radiotherapy. Results Glu and GABA are the key amino-acids in radio effection. After 12Gy or 24Gy irradiation there were low Glu, high GABA and low Glu/GABA in frontal lobe of rat. After 1h, 24h and 1 week irradiation, the contents of GABA raised up while the ratio of Glu/GABA declined with a remarkable decline at 24h. Conclusions Radiotherapy to epileptic rat may set off a rapid and enduring change of GABA and Glu transmitters, and 12Gy may be the rational radio dose for epileptic rat.

Objective To study the effects on inhibit tumor growth and enhance immune function of Ganoderma lucidum polysaccharide(GLP)combined with low-dose cisplatin in tumor-bearing mice.Methods 2 × 106 Lewis cells in 0.1 mL PBS were injected subcutaneously into the flanks of healthy female BALB/c nude mice (5～6 weeks old).When the tumor volume reached 100 mm3,mice were randomly assigned to 4 groups,control group (Oral gavage of 0.5 ml PBS and intraperitoneal injection of 0.5 ml),GLP group (gavage once every two days,40 mg/kg GLP in 0.5 ml),cisplatin group (intraperitoneal injection once every two days,2 mg/kg cisplatin in 0.5 ml) and the combination group (such as the above dose of cisplatin and GLP).After the treatment regimen was complete,the mice were sacrificed,and the tumors were removed,photographed,weighed,and prepared for histological analysis.The immune cells in spleen and peripheral blood were analyzed by flow cytometry.Results GLP can effectively increase anti-tumor effect of the small dose cisplatin,improve the survival of tumor-bearing mice,promote Th cells convert to Th1 in peripheral blood,increased the expression of CD 11c + on DC cell surface (GLP group 4.59％ VS the control group 1.06％,the combined group 7.21％ VS the cisplatin group 2.8％).Conclusion GLP can improve the immune function of tumor-bearing mice,had synergistic anti-tumor growth with low dose cisplatin,moreover,reduce renal toxic effects of cisplatin.

Objective To investigate the effects on self-renewal of pancreatic cancer stem cells by inhibiting hedgehog signaling pathway through cyclopamine. Methods PANC1 stem cells, PANC1 adherent cells and immortalized pancreatic ductal epithelial H6C7 cells were treated with 0.5, 1, 2, 5, 10 mol/L of cyclopamine for 24, 48, 72 h. The expression of Smo mRNA and Gli1 mRNA were detected by real-time PCR.Cell growth viability was measured by CCK 8. Cell cycle and apoptosis were determined by flow cytometry.Results Seventy-two hours after cyclopamine treatment, the Smo mRNA expressions of PANC1 stem cells,PANC1 adherent cells and H6C7 cells were 1,0.83 and 2.61; the expressions of Gli mRNA were 57.27,26.35,1; the inhibitory rates were ( 37.85 ± 13.69 ) %, ( 8.53 ± 4.43 ) %, (43.55 ± 28.98 ) %. Compared with PANC1, the expressions of Smo mRNA, Gli1 mRNA and the inhibitory rate of PANC1 stem cells significantly increased ( P ＜ 0.05 ). The proportion of G1 stage of PANC1 stem cells significantly decreased from (67.41 ±6.35)% to (36.53 ±6.03)% (P ＜0.05), and the apoptosis decreased from (10.95 ±5.68) % to ( 5.73 ± 1.42 ) % ( P ＞ 0.05 ). The proportion of G1 stage of PANC1 cells significantly decreased from ( 67.64 ± 6.88 ) % to ( 53.13 ± 1.10 ) % ( P ＜ 0.05 ); the apoptosis decreased from ( 12.08 ±4.12)% to (5.66 ± 1.33)% (P ＞0.05). While both the proportion of G1 stage and apoptosis of H6C7 cells was not significantly different. Conclusions Cyclopamine can inhibit the proliferation of PANC1 stem cells via blocking hedgehog signal pathway, and the mechanism may not be associated with cell apoptosis.

OBJECTIVETo compare steroidal saponins-main active ingredients of Paris polyphylla from different areas in Guizhou province, in order to provided basis for further proving Guizhou province to be the planting base of P. polyphylla.

METHODThe contents of nine steroidal saponins, namely paris-VII, (25R)-5-en-spirost-3beta,17alpha-diol-3-O-alpha-L-rhamnopyranosyl-(1-->2) [alpha-L-rhamnopyranosyl(1-->4)]-beta-D-glycopyanoside (PGRR), paris-H, paris-VI, paris-II , paris-III, gracillin, paris-I and paris-V of P. polyphylla from eight areas of Guizhou province were determined by HPLC-UV.

RESULTFive steroidal saponins-paris-VII, PGRR, paris-H, paris-VI and gracillin were detected in all the drugs, which provided basis for distinguishing P. polyphylla. P. polyphylla from Dushan showed the highest content of steroidal saponins (9.62%), followed by Tongren (6.39%), and the lowest was Zunyi (0.99%).

CONCLUSIONSaponins from different areas of Guizhou province show significant difference in variety and content, therefore Dushan is suitable for planting P. polyphylla.

China ; Chromatography, High Pressure Liquid ; methods ; Liliaceae ; chemistry ; Phytosterols ; analysis ; isolation & purification ; Plant Extracts ; analysis ; isolation & purification ; Rhizome ; chemistry ; Saponins ; analysis ; isolation & purification

Objective To establish the method of suspension culture for stem cells from human pancreatic cancer cell line PANC1.Methods PANC1 cells were cultured in serum-free medium under floating-culture system.Tumor cell spheres were observed by optical microscope.Expression of CD133 and cell cycle were detected by flow cytometry.Cancer stem cells were induced to differentiate with 10%FBS,and expression of CK18,was evaluated with immunofluorescence microscope.Spheres cells were injected into the subcutaneous space of NOD/SCID rat and tumor formation was monitored weekly.Results PANC1 cells could form the stem cells spheres,and the rate of sphere formation was stable between 4%0 and 5%0 after 20 passages in vitro.The expression of CD133(5.91±0.7%)and proportion of G0/G1 phase cell(80.99±2.60%)was significantly increased in spheres cells compared with parental PANC1 cells(1.44±0.52%and 69.01±5.03%),and the difference was statistically significant(P＜0.05).When these spheres cells were cultured in media with serum,these cells gradually returned to the status of parental cells and expressed CK18,2×103 sphere cells injection could initiate tumor fornmtion in NOD/SCID rat.Conclusions Tumor spheres stem cellscould be generated under serum-free floating-culture system.The sphere cells possessed the capacities of self renew,difierentiation,and tumorigenic potential.

Objective:To investigate the effect of rapamycin (Rapa) on apoptosis of acute myeloid leukemia THP-1 cells induced by idarubicin (IDA) and its molecular mechanism.Methods:The THP-1 cells were treated with 10, 20, 40 and 80 nmol/L Rapa for 1 h, and the cells without Rapa treatment were set up. Western blot was used to detect the conversion of autophagy marker LC3 protein in THP-1 cells (the ratio of LC3Ⅱ/LC3Ⅰ), flow cytometry was used to detect the apoptotic rate, and the pretreatment concentration of Rapa was determined. THP-1 cells were treated with different concentrations of IDA for 24 h, the cell proliferation inhibition rate of IDA for THP-1 cells was detected by CCK-8 method, and the half maximal inhibitory concentration ( IC50) was calculated. THP-1 cells with or without Rapa treatment were treated by IDA with the concentration of lower than IC50 for 24 h, CCK-8 method was used to detect cell proliferation inhibition rate, flow cytometry was used to detect cell apoptosis, real-time fluorescent quantitative polymerase chain reaction was used to detect the expression changes of autophagy-related genes Beclin-1, LC3 and p62, and Western blot was used to detect the conversion of autophagy marker LC3 protein. Results:The ratio of LC3Ⅱ/LC3Ⅰ in THP-1 cells treated by 20 nmol/L Rapa was higher than that in the untreated cells ( P=0.002 4). The apoptotic rate in THP-1 cells treated by 80 nmol/L Rapa was higher than that in the untreated cells ( P=0.007 3). According to the results of Western blot and flow cytometry, 20 nmol/L Rapa was selected as the pretreatment concentration. The IC50 of IDA for THP-1 cells treated with IDA for 24 h was 59.874 nmol/L. After treated with 50 nmol/L IDA for 24 h, the proliferation inhibitory [(69.67±5.03)% vs. (41.67±3.51)%] and apoptotic rates [(74.35±4.83)% vs. (41.25±5.24)%] in THP-1 cells pretreated by Rapa were higher than those in the unpretreated cells (both P<0.05); the Beclin-1 and LC3 mRNA expression levels and the ratio of LC3Ⅱ/LC3Ⅰ in THP-1 cells pretreated by Rapa were higher than those in the unpretreated cells, and the expression of p62 mRNA was lower than that in the unpretreated cells (all P<0.05). Conclusion:Rapa can enhance the apoptosis of THP-1 cells induced by a relative low dose of IDA, which may be achieved through inducing excessive autophagy in THP-1 cells.

BACKGROUND: Placenta mesenchymal stem cells have become hot spots in stem cells study in recent years because of its advantages. OBJECTIVE:To analyze the biological characteristics of amniotic mesenchymal stem cells and amnion epithelial cells, and to explore the feasibility of amniotic mesenchymal stem cells and amnion epithelial cells applied as seed cells in three-dimensional liquid culture to construct the tissue-engineered skin. METHODS:The amniotic mesenchymal stem cells and amnion epithelial cells were obtained by using multi-step digestion with trypsin and col agenase;then the flow cytometry, reverse transcription-PCR and immunofluorescent staining techniques were adopted to identify the surface molecular markers, stem cellcharacteristics and keratinocytes similarity respectively. Based on these data, amniotic mesenchymal stem cells and amnion epithelial cells used as seed cells together with rat type Ⅰ col agen matrix were adopted for three-dimensional liquid culture. RESULTS AND CONCLUSION:Flow cytometry test showed that amniotic mesenchymal stem cells and amnion epithelial cells cultured in vitro could highly express CD90, CD73 and CD105, and could not express the hemopoietic stem cellmarker of CD34 and MHC-class Ⅱ molecular HLA-DR. Reverse transcription-PCR results detected that amniotic mesenchymal stem cells could express stem cellcharacteristic genes CMCY and NANOG;amnion epithelial cells could express stem cellcharacteristic genes CMCY and KLF4, showing that both amniotic mesenchymal stem cells and amnion epithelial cells have stem cellproperties. Reverse transcription-PCR results showed that amniotic mesenchymal stem cells could express keratinocytes characteristic genes K19,β1-integrin and K8;amnion epithelial cells could express K19,β1-integrin, K5 and K8. Immunofluorescence staining results showed amnion epithelial cells could express keratinocytes proliferation related protein K14, which revealed that there was certain similarity in the mRNA expression between keratinocytes and amnion epithelial cells, and indicating that it has the potential to differentiate into keratinocytes. Tissue-engineered skin was successful y constructed by using amniotic mesenchymal stem cells and amnion epithelial cells, hematoxylin-eosin staining section showed that it has certain skin structure, and amnion epithelial cells had a preliminary differentiation. Al these prove that it is feasible to construct human skin tissues with amniotic mesenchymal stem cells and amniotic epithelial cells through the three-dimensional culture.

Objective To enhance the cognition about the adult contagious atypical pneumonia for an expectation of earlier diagnosis, prevention and treatment.Methods We analyzed and summarized X-ray lung and clinical characteristics of 34 patients with contagious atypical pneumonias who were admitted to our hospital from Feb to April. 2003.Results X-ray findings of lung were characterized by mostly involving unilateral and presenting ill defined cloud or patches-form opacities. These lesions progressed and varied rapidly. The clinical manifestations were characterized by the onset of fever (34/34), mostly associated with systemic and joint aching pains (29/34) and dry coughs (29/34). Leucocyte counts showed normal or lower than normal. Patient with a continuing high fever might tend to develop ARDS (2/34). All the 34 patients recovered after combined therapies (anti-inflammation, anti-virus and promoting immunity ) and were discharged from hospital. The average in-hospital days were (10.6?4.2) days. Conclusion X-ray examination is the main method for the early diagnosis and observing state-variation. The predominant clinical features of this disease are fever, dry cough, aching pain in extremities, unconspicuous pulmonary signs as well as hemogram not rising, a continuing high fever may lead to the tendency of ARDS.

We synthesized the superparamagnetic paclitaxel nanoparticles from modified chitosan tangling around Fe3O4 ferrofluid and taxol, and observed the nanoparticles with transmission electronic microscopy (TEM). Then we evaluated the paramagnetism of the particles by vibration specimen magnetometer (VSM) and tested their cytotoxicity with flow cytometry (FCM). The prepared nanoparticle solution was black without any floccule or sediment and appeared transparent after diluted. The nanoparticles were spherical and dispersed in water with mean diameter of 15 nm under TEM and showed superparamagnetic character. FCM test showed the nanoparticles had significant toxic effects against malignant astrocytoma U251 cell lines, equal to taxol alone. These results showed that the superparamagnetic nanoparticle not only enhanced the solubility of paclitaxel in water, but also was superparamagnetic and cytotoxic, which make suitable tools for magnetic targeting chemotherapy of brain gliomas.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Brain Neoplasms ; pathology ; Cell Line, Tumor ; Chitosan ; pharmacology ; Drug Carriers ; chemistry ; Drug Compounding ; methods ; Ferric Compounds ; Glioma ; pathology ; Humans ; Magnetics ; Metal Nanoparticles ; chemistry ; Nanoparticles ; Paclitaxel ; pharmacology

Objective:To investigate the treatment and prognosis of children with propionic acidemia (PA).Methods:This study involved 82 children with PA treated in the Department of Pediatric Endocrinol-ogy and Genetic Metabolism, Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from December 2002 to June 2020. Clinical data, including manifestations, laboratory test results, treatment strategy, and follow-up data, were summarized and analyzed using t-test or Mann-Whitney U test. Results:(1) Among the 82 cases consisting of 50 (61.0%) boys and 32 (39.0%) girls, 59 (72.0%) were diagnosed after clinical onset; 22 (26.8%) were diagnosed by newborn screening, including eight asymptomatic ones; the other one (1.2%) was asymptomatic but confirmed after the diagnosis of PA in the patient's sibling. The average age at first onset was 4.5 months (2 d-5 years) in 73 subjects, of which 28 (38.4%) were early-onset PA (within three months after birth). (2) Cranial MRI was performed on 26 cases, and abnormality was identified in 19 (73.1%) cases. (3) Hyperlactatemia was found in 16 cases among 30(53.3%) who underwent relevant examination with the average lactic acid level of 3.5 (2.1-4.3) μmol/L, while 35 out of 40 patients (87.5%) had hyperammonemia with an average blood ammonia level of 105.4 (34-907) μmol/L. (4) Among the 28 early-onset PA cases, 16 (57.1%) died, and 12 (42.9%) survived. There was no significant difference in the serum propionylcarnitine level, propionylcarnitine to acetylcarnitine ratio, urine 3-hydroxypropionic acid, or methylcitrate level between the survival and death cases. (5) Genetic mutations were detected in 75 patients (91.5%), among which 26 (34.7%) carried PCCA gene mutations and 48 (64%) with PCCB gene mutations. One patient (1.3%) harbored one known pathogenic mutation in each of the PCCA and PCCB genes. All mutations were inherited from the parents. (6) Followed up to June 2020, 57 (69.5%) patients survived, and 25 (30.5%) died from multiple organ failure secondary to severe acidosis, including 16 early-onset and nine late-onset cases. Conclusions:The primary treatment of PA is dietary control. Most PA patients are diagnosed after clinical onset, but symptoms may recur and even have developmental retardation despite treatment. Some of those diagnosed through newborn screening are asymptomatic after treatment. Newborn screening using tandem mass spectrometry is recommended for early diagnosis and treatment of PA.